CN106070185B - One kind is for fat marrow acquisition suit - Google Patents
One kind is for fat marrow acquisition suit Download PDFInfo
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- CN106070185B CN106070185B CN201610622783.6A CN201610622783A CN106070185B CN 106070185 B CN106070185 B CN 106070185B CN 201610622783 A CN201610622783 A CN 201610622783A CN 106070185 B CN106070185 B CN 106070185B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
- A61N1/00—Electrotherapy; Circuits therefor
- A61N1/02—Details
Abstract
The present invention provides a kind of for fat marrow acquisition suit, including collecting cassette, it is provided with the protection liquid bottle of fat marrow Cell protective solutions, two collection tubes, several ice chests, acquisition suit operation instruction, capture card and a set of bar code, the collecting cassette includes box body, hinged box cover and the collet of bottom in box body is mounted on the box body, the collet is equipped with the protection liquid bottle jack for placing protection liquid bottle and two collection tube jacks for placing collection tube, the partition around the collet is equipped in the box body, the partition is vertical with box bottom, the ice chest is placed in the cavity formed between the box side wall and the partition, the box cover upper surface is additionally provided with for placing acquisition suit operation instruction, the groove of capture card and bar code, the groove is equipped with the cover board with matching grooves.The present invention has the function of heat-insulated, damping, temperature monitoring, can effectively ensure that the storage and transport safety of the fat marrow cell of acquisition.
Description
Technical field
The invention belongs to the technical fields that cell saves, more particularly to a kind of acquire for fat marrow to use suit.
Background technique
In recent years, cell technology is widely used in the clinical test and treatment of a variety of diseases, and shows good
Application prospect is increasingly becoming one of medical research hot spot new in recent years.High-quality cell is acquired from healthy human body to be saved,
It can be used for disease treatment, damage maintenance, anti-aging and sub-health treatment etc., play treatment disease or even save patient vitals
Effect.To guarantee that the cell of acquisition plays a role well in the therapeutic process in later period, the acquisition and preservation process of cell
Particularly significant, the preservation medium frequently with physiological saline or cell culture medium as cell, is maintaining cell activity, is protecting at present
Demonstrate,prove cell survival rate in terms of it is ineffective, and save process safety and reliability it is also not ideal enough；Existing preservation
The influences of the factors to cell survival rate and stability such as the container of cell does not fully take into account temperature, transport is jolted, lead to cell
The effect of preservation is not ideal enough.
Summary of the invention
It uses and is set with for fat marrow acquisition in order to solve the above technical problem, the present invention provides one kind, including collecting cassette,
It is provided with the protection liquid bottle, two collection tubes, several ice chests, acquisition suit operation instruction, acquisition of fat marrow Cell protective solutions
Card and a set of bar code, the collecting cassette include box body and the hinged box cover of the box body and the bottom for being mounted on bottom in box body
Support, the collet is equipped with the protection liquid bottle jack for placing protection liquid bottle and two collection tubes for placing collection tube are inserted
Hole, the interior partition being equipped with around the collet of the box body, the partition is vertical with box bottom, and the ice chest is placed on described
In the cavity formed between box side wall and the partition, the box cover upper surface is additionally provided with to be said for placing to acquire to be set with to use
The groove of bright, capture card and bar code, the groove are equipped with the cover board with matching grooves.
Further, flexible fastening circle is equipped on the protection liquid bottle jack and collection tube jack.
Further, the partition upper end is equipped with inner cover, and the inner surface of the inner cover is equipped with and the protection liquid bottle
The protection liquid bottle cap hole matched and with the matched collection tube cap bore of the collection tube, play and protection liquid bottle and collection tube be auxiliarily fixed
Effect, prevent protection liquid bottle and collection tube run-off the straight or shaking.
Further, device for monitoring temperature is additionally provided on the collecting cassette.
Further, the acquisition box outer surfac is equipped with anticollision buffer layer.
Further, insulating layer is equipped between the acquisition box outer surfac and anticollision buffer layer.
Further, the box body bottom is equipped with anti-skid device.It is conventional that security bar, sucker etc. can be used in the anti-skid device
Anti-skidding means.
Further, the junction of the box body and box cover is equipped with fixed device.The fixed device can using buckle,
Lock collar or ligature play the role of that box cover is prevented to be opened accidentally.
Further, the fat marrow Cell protective solutions contained in the protection liquid bottle are mainly by hypoxemia protective agent, dimension
Raw element C and aminoglycoside antibiotics are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium described in every mL
Aqueous solution dissolves the hypoxemia protective agent 5-10mg, vitamin C 10-50 μ g, the aminoglycoside antibiotics respectively
50-200U, the concentration of the DMEM/F12 culture medium aqueous solution are 30-40mg/mL.
DMEM/F12 culture medium is combined by the ratio that DMEM culture medium and F12 culture medium are 1:1 according to parts by weight, at
It is point abundant, nutrient concentration is high, contains various trace elements, can be used as serum-free or serum content it is lower under the conditions of mammal
The basal medium of cell culture.
Contain following ingredients in every liter of DMEM/F12 culture medium:
Hypoxemia protective agent, vitamin C and ammonia are added in the DMEM/F12 culture medium aqueous solution that concentration is 30-40mg/mL
Base glycoside antibiotic, fat marrow Cell protective solutions obtained can keep fat stem cell or medulla mesenchyma dry for a long time
Cell activity improves the survival rate of fat stem cell or mesenchymal stem cell.
Further, the aminoglycoside antibiotics is by one of gentamicin, kanamycins, streptomysin or a variety of
Composition.
Preferably, the aminoglycoside antibiotics by gentamicin, kanamycins, streptomysin according to 2:2:1 weight
Portion rate composition, than a kind of aminoglycoside antibiotics is applied alone, using other several aminoglycoside antibiotics or using other
The anti-bacterial effect of weight proportion is more preferable.
Further, benzalkonium chloride and calcium formate, DMEM/F12 culture medium water described in every mL are also contained in the protection liquid
Solution dissolves benzalkonium chloride 50-100 μ g, the calcium formate 50-100 μ g respectively.Protection liquid in be added benzalkonium chloride and
Calcium formate plays the role of preventing generating fungi and virus infection in fat marrow Cell protective solutions, ensures saved cell quality
Safety, can be applied to the treatment of human body diseases.
Further, the hypoxemia protective agent includes each ingredient of following parts by weight:
Banaba leaf extract 2-5 Herba Oxalidis Corniculatae extract 2-5 oxymatrine 1-2.
Made in fat marrow Cell protective solutions using banaba leaf extract, Herba Oxalidis Corniculatae extract and oxymatrine
Reactive oxygen species in fat stem cell or mesenchymal stem cell can be significantly reduced for hypoxemia protective agent, it is dry thin to adjust fat
Born of the same parents or mesenchymal stem cell oxidation resistance reduce withering for fat stem cell caused by hypoxemia or mesenchymal stem cell
It dies, effectively improves the survival rate of the fat stem cell or mesenchymal stem cell under hypoxia condition.
Further, the hypoxemia protective agent further includes each ingredient of following parts by weight:
Erythrothioneine 2-5 dibutyl hydroxy toluene 1-3.
Erythrothioneine is added in hypoxemia protective agent and dibutyl hydroxy toluene is mentioned with banaba leaf extract, creeping oxalis
It takes object and oxymatrine to be used cooperatively, the survival of the fat stem cell or mesenchymal stem cell under hypoxia condition can be made
Rate is higher.
Further, hyaluronic acid, chitin and Heparan sulfate are also contained in the protection liquid, described in every mL
DMEM/F12 culture medium aqueous solution dissolves the hyaluronic acid 10-20mg, the chitin 1-5mg, the acetyl sulfate respectively
Heparin 1-5mg.
Hyaluronic acid, chitin and Heparan sulfate is added in protection liquid, can effectively improve fat stem cell or
The survival rate of mesenchymal stem cell.
Further, trehalose, hydroxypropyl cyclodextrin and sodium alginate are also contained in the protection liquid, described in every mL
DMEM/F12 culture medium aqueous solution dissolves the trehalose 1-5mg, hydroxypropyl cyclodextrin 10-50 μ g, the seaweed respectively
Sour sodium 10-50 μ g.
Trehalose, hydroxypropyl cyclodextrin and sodium alginate is added in protection liquid, can play improve fat stem cell or
The effect of the growth rate of mesenchymal stem cell.
Further, glucan, hypromellose and lauric monoglyceride, every mL are also contained in the protection liquid
The DMEM/F12 culture medium aqueous solution dissolves the glucan 3-5mg, the hypromellose 3-5mg, described respectively
Lauric monoglyceride 50-100 μ g.
Glucan, hypromellose and lauric monoglyceride is added in protection liquid, fat can be effectively improved
Bone marrow cell protects the stability of liquid, extends the shelf-life of fat marrow Cell protective solutions.
Further, the preparation method with the fat marrow Cell protective solutions in suit is acquired for fat marrow, comprising:
(1) NaHCO is added into DMEM/F12 culture medium aqueous solution3, until the pH value of DMEM/F12 culture medium aqueous solution
For 6-6.5；
(2) other raw materials in addition to vitamin C are added in the DMEM/F12 culture medium aqueous solution that step (1) obtains,
Gained mixed liquor is warming up to 80-100 DEG C again, stirring is to being uniformly dissolved；
(3) mixed liquor that step (2) obtains is cooled to 20-30 DEG C, is added vitamin C, stirring is to being uniformly dissolved；
(4) mixed liquor that step (3) obtains is filtered with biofilter, is then cooled to 0-10 DEG C in ice-water bath
Obtain fat marrow Cell protective solutions.
Further, the step (2) carries out under conditions of pressure is 0.16-0.2MPa.
It is stirred operation under pressurized constraint, the effect of stirring can be improved, makes in fat marrow Cell protective solutions
Various raw materials are sufficiently mixed uniformly, improve the stability of fat marrow Cell protective solutions.
The present invention can be used for acquisition, the storage and transport of fat or bone marrow cell, with heat-insulated, damping, temperature monitoring
Function can place multiple ice chests to maintain the stabilization of temperature, and the fat of acquisition and the storage and transport of bone marrow cell are effectively ensured
Safety；Fat marrow Cell protective solutions performance provided by the invention is stablized, and can save fat cell or marrow for a long time
Mescenchymal stem cell, play and effectively keep the activity of fat stem cell or mesenchymal stem cell, improve fat stem cell or
The effect of the proliferative capacity of mesenchymal stem cell.
Detailed description of the invention
Fig. 1 is the structural schematic diagram for fat marrow acquisition suit of the embodiment of the present invention 1；
Fig. 2 is the perspective view for fat marrow acquisition suit of the embodiment of the present invention 1；
Fig. 3 is the structural schematic diagram for fat marrow acquisition suit of the embodiment of the present invention 2；
Fig. 4 is the structural schematic diagram for fat marrow acquisition suit of the embodiment of the present invention 3；
Fig. 5 is the perspective view for fat marrow acquisition suit of the embodiment of the present invention 4；
Fig. 6 is the bottom view for fat marrow acquisition suit of the embodiment of the present invention 4；
Fig. 7 is the fat stem cell growth curve chart of the embodiment of the present invention 8, embodiment 23, reference examples 13 and reference examples 14；
Wherein: 1 collecting cassette, 2 protection liquid bottles, 3 collection tubes, 4 ice chests, 101 box bodys, 102 box covers, 103 collets, 104 protections
Liquid bottle jack, 105 collection tube jacks, 106 partitions, 107 grooves, 108 cover boards, 109 flexible fastening circles, 110 inner covers, 111 protections
Liquid bottle cap hole, 112 collection tube cap bore, 113 anti-skid devices, 114 fixed devices, 115 attackers.
Specific embodiment
Embodiment 1
One kind as depicted in figs. 1 and 2, including collecting cassette 1, being provided with fat marrow for fat marrow acquisition suit
2, two collection tubes 3 of protection liquid bottle, several ice chests 4, acquisition suit operation instruction, capture card and a set of bar shaped of Cell protective solutions
Code, the collecting cassette 1 include box body 101 and the hinged box cover 102 of the box body and the collet for being mounted on bottom in box body 101
103, the collet 103 is equipped with the protection liquid bottle jack 104 and two for placing collection tube 3 for placing protection liquid bottle 2
A collection tube jack 105, the interior partition 106 being equipped with around the collet 103 of the box body 101, the partition 106 and box body
101 plane perpendiculars, the ice chest 4 are placed in the cavity formed between 1 side wall of box body and the partition 106, the box
102 upper surface of lid is additionally provided with the groove 107 for placing acquisition suit operation instruction, capture card and bar code, the groove 107
It is equipped with and the matched cover board 108 of groove 107.
Before carrying out cell collection, operator is first turned on the cover board 108 of 102 upper surface of box cover, takes out from groove 107
Acquisition suit operation instruction, capture card and bar code read over acquisition suit operation instruction, and capture card and bar code is quasi-
It gets ready spare, then opens the lid 102, take out protection liquid bottle 2 inside box body 101 and two collection tubes 3 are spare；Acquisition is completed
The fat of acquisition or marrow are put into collection tube 3 afterwards, every pipe 25ml, then protect 25ml bone marrow cell from protection liquid bottle 2
Shield liquid pours into collection tube 3, tightens the cap of collection tube 3, and bar code is attached on collection tube 3, then collection tube 3 is vertically put
Enter the collection tube jack 105 in box body 101, then the ice chest 4 frozen is put between 101 side wall of box body and partition 106 again
It in cavity and covers on box cover 102, guarantees that the temperature in collecting cassette meets cell and saves requirement, finally filled out in detail on capture card
Donor data and acquisition information are write, and sticks bar code, is put back in groove 107.
Embodiment 2
One kind is for fat marrow acquisition suit, as shown in figure 3, unlike the first embodiment: the protection liquid bottle is inserted
Flexible fastening circle 109, the cavity between 1 side wall of box body and the partition 106 are equipped on hole 104 and collection tube jack 105
4 ice chest placing grooves are divided by 4 pieces of risers perpendicular to 101 bottom surface of box body.
Embodiment 3
One kind is for fat marrow acquisition suit, as shown in figure 4, unlike the first embodiment: on the partition 106
End is equipped with inner cover 110, the inner surface of the inner cover 110 be equipped with the matched protection liquid bottle cap hole 111 of the protection liquid bottle 2 with
And with the matched collection tube cap bore 112 of the collection tube 3, device for monitoring temperature, the temperature prison are additionally provided on the collecting cassette 1
Control device includes temperature measurer inside the box body 101 and processor and display screen and loudspeaker positioned at 101 surface of box body,
The processor is connected respectively at temperature measurer, display screen with loudspeaker.
Embodiment 4
One kind is for fat marrow acquisition suit, as shown in Figure 5 and Figure 6, unlike the first embodiment: the acquisition
1 outer surface of box is successively arranged insulating layer and anticollision buffer layer from inside to outside, and 101 bottom of box body is equipped with anti-skid device 113, institute
Stating anti-skid device 113 is that several are mounted on the sucker of 101 bottom of box body, and the junction of the box body 101 and box cover 102 is equipped with
Fixed device 114, the fixed device 114 are buckle, and the cover board 108 is equipped with attacker 115.
Embodiment 5
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
It is mainly that hypoxemia protective agent, vitamin C and gentamicin is molten with DMEM/F12 culture medium aqueous solution that fat bone marrow cell, which protects liquid,
It solves, DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 8mg, 20 μ of the vitamin C respectively
G, the gentamicin 100U, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 6
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
It is mainly that hypoxemia protective agent, vitamin C and gentamicin is molten with DMEM/F12 culture medium aqueous solution that fat bone marrow cell, which protects liquid,
It solves, DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 5mg, 10 μ of the vitamin C respectively
G, the gentamicin 50U, the concentration of the DMEM/F12 culture medium aqueous solution are 30mg/mL.
Embodiment 7
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
It is mainly that hypoxemia protective agent, vitamin C and gentamicin is molten with DMEM/F12 culture medium aqueous solution that fat bone marrow cell, which protects liquid,
It solves, DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 10mg, 50 μ of the vitamin C respectively
G, the gentamicin 200U, the concentration of the DMEM/F12 culture medium aqueous solution are 40mg/mL.
Embodiment 8
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin DMEM/F12
Culture medium aqueous dissolution forms, and DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 8mg, institute respectively
State 20 μ g of vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the DMEM/F12 culture
The concentration of base aqueous solution is 37.5mg/mL.
Embodiment 9
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin DMEM/F12
Culture medium aqueous dissolution forms, and DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 5mg, institute respectively
State 10 μ g of vitamin C, the gentamicin 20U, the kanamycins 20U, the streptomysin 10U, the DMEM/F12 culture
The concentration of base aqueous solution is 30mg/mL；
The preparation method of the fat marrow Cell protective solutions includes:
(1) NaHCO3 is added into DMEM/F12 culture medium aqueous solution, until the pH value of DMEM/F12 culture medium aqueous solution
It is 6；
(2) other raw materials in addition to vitamin C are added in the DMEM/F12 culture medium aqueous solution that step (1) obtains,
Gained mixed liquor is warming up to 80 DEG C again, stirring is to being uniformly dissolved；
(3) mixed liquor that step (2) obtains is cooled to 20 DEG C, is added vitamin C, stirring is to being uniformly dissolved；
(4) mixed liquor that step (3) obtains is filtered with biofilter, 0 DEG C is then cooled in ice-water bath to obtain
Obtain fat marrow Cell protective solutions.
Embodiment 10
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly to train hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin DMEM/F12
It supports base aqueous dissolution to form, DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 10mg, institute respectively
State 50 μ g of vitamin C, the gentamicin 80U, the kanamycins 80U, the streptomysin 40U, the DMEM/F12 culture
The concentration of base aqueous solution is 40mg/mL.
Embodiment 11
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride,
Calcium formate is formed with DMEM/F12 culture medium aqueous dissolution, described in DMEM/F12 culture medium aqueous solution dissolves respectively described in every mL
Hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U,
75 μ g of benzalkonium chloride, 75 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 12
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride,
Calcium formate is formed with DMEM/F12 culture medium aqueous dissolution, described in DMEM/F12 culture medium aqueous solution dissolves respectively described in every mL
Hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U,
50 μ g of benzalkonium chloride, 50 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL；
The preparation method of the fat marrow Cell protective solutions includes:
(1) NaHCO3 is added into DMEM/F12 culture medium aqueous solution, until the pH value of DMEM/F12 culture medium aqueous solution
It is 6.5；
(2) other raw materials in addition to vitamin C are added in the DMEM/F12 culture medium aqueous solution that step (1) obtains,
Gained mixed liquor is warming up to 100 DEG C again, stirring is to being uniformly dissolved；
(3) mixed liquor that step (2) obtains is cooled to 30 DEG C, is added vitamin C, stirring is to being uniformly dissolved；
(4) mixed liquor that step (3) obtains is filtered with biofilter, 10 DEG C are then cooled in ice-water bath to obtain
Obtain fat marrow Cell protective solutions.
Embodiment 13
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride,
Calcium formate is formed with DMEM/F12 culture medium aqueous dissolution, described in DMEM/F12 culture medium aqueous solution dissolves respectively described in every mL
Hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U,
100 μ g of benzalkonium chloride, 100 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 14
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly that banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, vitamin C, celebrating is big
Mycin, kanamycins, streptomysin are formed with DMEM/F12 culture medium aqueous dissolution, and DMEM/F12 culture medium described in every mL is water-soluble
Liquid dissolve respectively the banaba leaf extract 3.5mg, the Herba Oxalidis Corniculatae extract 3mg, the oxymatrine 1.5mg,
20 μ g of vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the DMEM/F12 training
The concentration for supporting base aqueous solution is 37.5mg/mL.
Embodiment 15
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly that banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, vitamin C, celebrating is big
Mycin, kanamycins, streptomysin, benzalkonium chloride, calcium formate are formed with DMEM/F12 culture medium aqueous dissolution, described in every mL
DMEM/F12 culture medium aqueous solution dissolve respectively the banaba leaf extract 3.2mg, the Herba Oxalidis Corniculatae extract 3.2mg,
The oxymatrine 1.6mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the strepto-
Plain 20U, 75 μ g of the benzalkonium chloride, 75 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/
mL；
The preparation method of the fat marrow Cell protective solutions includes:
(1) NaHCO3 is added into DMEM/F12 culture medium aqueous solution, until the pH value of DMEM/F12 culture medium aqueous solution
It is 6.3；
(2) other raw materials in addition to vitamin C are added in the DMEM/F12 culture medium aqueous solution that step (1) obtains,
Gained mixed liquor is warming up to 90 DEG C again, stirring is to being uniformly dissolved；
(3) mixed liquor that step (2) obtains is cooled to 23 DEG C, is added vitamin C, stirring is to being uniformly dissolved；
(4) mixed liquor that step (3) obtains is filtered with biofilter, 4 DEG C are then cooled in ice-water bath to obtain
Obtain fat marrow Cell protective solutions.
Embodiment 16
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly that banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, vitamin C, celebrating is big
Mycin, kanamycins, streptomysin, benzalkonium chloride, calcium formate are formed with DMEM/F12 culture medium aqueous dissolution, described in every mL
DMEM/F12 culture medium aqueous solution dissolves the banaba leaf extract 3.33mg, the Herba Oxalidis Corniculatae extract respectively
3.33mg, the oxymatrine 1.34mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U,
The streptomysin 20U, 75 μ g of the benzalkonium chloride, 75 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution
For 37.5mg/mL.
Embodiment 17
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin are formed with DMEM/F12 culture medium aqueous dissolution,
DMEM/F12 culture medium aqueous solution described in every mL dissolves the banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract respectively
2mg, the oxymatrine 1mg, the erythrothioneine 2mg, the dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C,
The gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the concentration of the DMEM/F12 culture medium aqueous solution
For 37.5mg/mL.
Embodiment 18
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate DMEM/F12 culture medium
Aqueous dissolution forms, DMEM/F12 culture medium aqueous solution described in every mL dissolve respectively the banaba leaf extract 1.8mg,
The Herba Oxalidis Corniculatae extract 1.8mg, the oxymatrine 1mg, the erythrothioneine 2.2mg, the dibutyl hydroxy toluene
1.2mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the benzene are pricked
75 μ g of oronain, 75 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL；
The preparation method of the fat marrow Cell protective solutions includes:
(1) NaHCO3 is added into DMEM/F12 culture medium aqueous solution, until the pH value of DMEM/F12 culture medium aqueous solution
It is 6.3；
(2) other raw materials in addition to vitamin C are added in the DMEM/F12 culture medium aqueous solution that step (1) obtains,
Gained mixed liquor is warming up to 90 DEG C again, is stirred in the case where pressure is 0.2MPa to being uniformly dissolved；
(3) mixed liquor that step (2) obtains is cooled to 23 DEG C, is added vitamin C, stirring is to being uniformly dissolved；
(4) mixed liquor that step (3) obtains is filtered with biofilter, 4 DEG C are then cooled in ice-water bath to obtain
Obtain fat marrow Cell protective solutions.
Embodiment 19
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate DMEM/F12 culture medium
Aqueous dissolution forms, and DMEM/F12 culture medium aqueous solution described in every mL dissolves the banaba leaf extract 2mg, institute respectively
State Herba Oxalidis Corniculatae extract 2mg, the oxymatrine 0.8mg, the erythrothioneine 2mg, the dibutyl hydroxy toluene
1.2mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the benzene are pricked
75 μ g of oronain, 75 μ g of the calcium formate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 20
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, hyaluronic acid,
Chitin, Heparan sulfate are formed with DMEM/F12 culture medium aqueous dissolution, and DMEM/F12 culture medium described in every mL is water-soluble
Liquid dissolve respectively the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U,
The streptomysin 20U, the hyaluronic acid 15mg, the chitin 2mg, the Heparan sulfate 2mg, the DMEM/F12
The concentration of culture medium aqueous solution is 37.5mg/mL.
Embodiment 21
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate, hyaluronic acid, crust
Matter, Heparan sulfate are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium aqueous solution described in every mL point
The banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract 2mg, the oxymatrine 1mg, the ergot are not dissolved
Thioneine 2mg, the dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins
40U, the streptomysin 20U, 75 μ g of the benzalkonium chloride, 75 μ g of the calcium formate, the hyaluronic acid 10mg, the chitin
1mg, the Heparan sulfate 1mg, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL；
The preparation method of the fat marrow Cell protective solutions includes:
(1) NaHCO3 is added into DMEM/F12 culture medium aqueous solution, until the pH value of DMEM/F12 culture medium aqueous solution
It is 6.3；
(2) other raw materials in addition to vitamin C are added in the DMEM/F12 culture medium aqueous solution that step (1) obtains,
Gained mixed liquor is warming up to 90 DEG C again, is stirred in the case where pressure is 0.18MPa to being uniformly dissolved；
(3) mixed liquor that step (2) obtains is cooled to 23 DEG C, is added vitamin C, stirring is to being uniformly dissolved；
(4) mixed liquor that step (3) obtains is filtered with biofilter, 4 DEG C are then cooled in ice-water bath to obtain
Obtain fat marrow Cell protective solutions.
Embodiment 22
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate, hyaluronic acid, crust
Matter, Heparan sulfate are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium aqueous solution described in every mL point
The banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract 2mg, the oxymatrine 1mg, the ergot are not dissolved
Thioneine 2mg, the dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins
40U, the streptomysin 20U, 75 μ g of the benzalkonium chloride, 75 μ g of the calcium formate, the hyaluronic acid 20mg, the chitin
5mg, the Heparan sulfate 5mg, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 23
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, trehalose, hydroxyl
Propyl cyclodextrin, sodium alginate are formed with DMEM/F12 culture medium aqueous dissolution, and DMEM/F12 culture medium described in every mL is water-soluble
Liquid dissolve respectively the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U,
The streptomysin 20U, the trehalose 2mg, 30 μ g of the hydroxypropyl cyclodextrin, the sodium alginate 30 μ g, the DMEM/
The concentration of F12 culture medium aqueous solution is 37.5mg/mL.
Embodiment 24
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate, hyaluronic acid, crust
Matter, Heparan sulfate, trehalose, hydroxypropyl cyclodextrin, sodium alginate are formed with DMEM/F12 culture medium aqueous dissolution, often
DMEM/F12 culture medium aqueous solution described in mL dissolves the banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract respectively
2mg, the oxymatrine 1mg, the erythrothioneine 2mg, the dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C,
The gentamicin 40U, the kanamycins 40U, the streptomysin 20U, 75 μ g of the benzalkonium chloride, 75 μ of the calcium formate
G, the hyaluronic acid 15mg, the chitin 2mg, the Heparan sulfate 2mg, the trehalose 1mg, the hydroxypropyl
10 μ g of cyclodextrin, 10 μ g of the sodium alginate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 25
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate, hyaluronic acid, crust
Matter, Heparan sulfate, trehalose, hydroxypropyl cyclodextrin, sodium alginate are formed with DMEM/F12 culture medium aqueous dissolution, often
DMEM/F12 culture medium aqueous solution described in mL dissolves the banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract respectively
2mg, the oxymatrine 1mg, the erythrothioneine 2mg, the dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C,
The gentamicin 40U, the kanamycins 40U, the streptomysin 20U, 75 μ g of the benzalkonium chloride, 75 μ of the calcium formate
G, the hyaluronic acid 15mg, the chitin 2mg, the Heparan sulfate 2mg, the trehalose 5mg, the hydroxypropyl
50 μ g of cyclodextrin, 50 μ g of the sodium alginate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 26
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, glucan, hydroxyl
Third methylcellulose, lauric monoglyceride are formed with DMEM/F12 culture medium aqueous dissolution, the training of DMEM/F12 described in every mL
Support base aqueous solution dissolve respectively the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, it is described card that
Mycin 40U, the streptomysin 20U, the bextran 45 mg, the hypromellose 4mg, the lauric monoglyceride
75 μ g, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Embodiment 27
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate, hyaluronic acid, crust
Matter, Heparan sulfate, trehalose, hydroxypropyl cyclodextrin, sodium alginate, glucan, hypromellose, lauric acid list are sweet
Grease is formed with DMEM/F12 culture medium aqueous dissolution, and DMEM/F12 culture medium aqueous solution described in every mL dissolves described big respectively
It is flower banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract 2mg, the oxymatrine 1mg, the erythrothioneine 2mg, described
Dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin
20U, 75 μ g of the benzalkonium chloride, 75 μ g of the calcium formate, the hyaluronic acid 15mg, the chitin 2mg, the sulfuric acid second
Acyl heparin 2mg, the trehalose 2mg, 30 μ g of the hydroxypropyl cyclodextrin, 30 μ g of the sodium alginate, the glucan 3mg,
The hypromellose 3mg, 50 μ g of the lauric monoglyceride, the concentration of the DMEM/F12 culture medium aqueous solution are
37.5mg/mL。
Embodiment 28
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, two
Butylated hydroxytoluene, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride, calcium formate, hyaluronic acid, crust
Matter, Heparan sulfate, trehalose, hydroxypropyl cyclodextrin, sodium alginate, glucan, hypromellose, lauric acid list are sweet
Grease is formed with DMEM/F12 culture medium aqueous dissolution, and DMEM/F12 culture medium aqueous solution described in every mL dissolves described big respectively
It is flower banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract 2mg, the oxymatrine 1mg, the erythrothioneine 2mg, described
Dibutyl hydroxy toluene 1mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin
20U, 75 μ g of the benzalkonium chloride, 75 μ g of the calcium formate, the hyaluronic acid 15mg, the chitin 2mg, the sulfuric acid second
Acyl heparin 2mg, the trehalose 2mg, 30 μ g of the hydroxypropyl cyclodextrin, 30 μ g of the sodium alginate, the glucan 5mg,
The hypromellose 5mg, 100 μ g of the lauric monoglyceride, the concentration of the DMEM/F12 culture medium aqueous solution
For 37.5mg/mL.
Reference examples 1
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protection liquid mainly forms hypoxemia protective agent and gentamicin with DMEM/F12 culture medium aqueous dissolution, often
DMEM/F12 culture medium aqueous solution described in mL dissolves the hypoxemia protective agent 8mg, the gentamicin 100U respectively, described
The concentration of DMEM/F12 culture medium aqueous solution is 37.5mg/mL.
Reference examples 2
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, cystine and gentamicin DMEM/F12 culture medium aqueous dissolution
It forms, DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 8mg, 20 μ g of the cystine, institute respectively
Gentamicin 100U is stated, the concentration of the DMEM/F12 culture medium aqueous solution is 37.5mg/mL.
Reference examples 3
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, penicillin with DMEM/F12 culture medium aqueous dissolution and
At DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, institute respectively
Penicillin 100U is stated, the concentration of the DMEM/F12 culture medium aqueous solution is 37.5mg/mL.
Reference examples 4
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin DMEM/F12
Culture medium aqueous dissolution forms, and DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 8mg, institute respectively
State 20 μ g of vitamin C, the gentamicin 20U, the kanamycins 60U, the streptomysin 20U, the DMEM/F12 culture
The concentration of base aqueous solution is 37.5mg/mL.
Reference examples 5
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride
It is formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia respectively to be protected
Protect agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the benzene
75 μ g of oronain is pricked, the concentration of the DMEM/F12 culture medium aqueous solution is 37.5mg/mL.
Reference examples 6
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, benzalkonium chloride,
Potassium sulfate is formed with DMEM/F12 culture medium aqueous dissolution, described in DMEM/F12 culture medium aqueous solution dissolves respectively described in every mL
Hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U,
75 μ g of benzalkonium chloride, 75 μ g of the potassium sulfate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Reference examples 7
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
It is mainly that vitamin C, gentamicin, kanamycins, streptomysin is water-soluble with DMEM/F12 culture medium that fat bone marrow cell, which protects liquid,
Liquid dissolves, and DMEM/F12 culture medium aqueous solution described in every mL dissolves 20 μ g of vitamin C, the gentamicin respectively
40U, the kanamycins 40U, the streptomysin 20U, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Reference examples 8
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly that isatis root extract, Herba Oxalidis Corniculatae extract, oxymatrine, vitamin C, celebrating is big mould
Element, kanamycins, streptomysin are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium aqueous solution described in every mL
The isatis root extract 3.5mg, the Herba Oxalidis Corniculatae extract 3mg, the oxymatrine 1.5mg, the dimension are dissolved respectively
Raw 20 μ g of element C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U, the DMEM/F12 culture medium water
The concentration of solution is 37.5mg/mL.
Reference examples 9
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, dimension
Raw element C, gentamicin, kanamycins, streptomysin are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 described in every mL
Culture medium aqueous solution dissolves the banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract 2mg, the Oxymatrine respectively
Alkali 1mg, the erythrothioneine 3mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the chain
Mycin 20U, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Reference examples 10
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid to be mainly by banaba leaf extract, Herba Oxalidis Corniculatae extract, oxymatrine, erythrothioneine, resist
Bad hematic acid sodium, vitamin C, gentamicin, kanamycins, streptomysin are formed with DMEM/F12 culture medium aqueous dissolution, every mL
The DMEM/F12 culture medium aqueous solution dissolve respectively the banaba leaf extract 2mg, the Herba Oxalidis Corniculatae extract 2mg,
The oxymatrine 1mg, the erythrothioneine 2mg, the sodium ascorbate 1mg, 20 μ g of the vitamin C, the celebrating are big
Mycin 40U, the kanamycins 40U, the streptomysin 20U, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/
mL。
Reference examples 11
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, hyaluronic acid,
Heparan sulfate is formed with DMEM/F12 culture medium aqueous dissolution, and the difference of DMEM/F12 culture medium aqueous solution described in every mL is molten
Solve the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the strepto-
Plain 20U, the hyaluronic acid 15mg, the Heparan sulfate 2mg, the concentration of the DMEM/F12 culture medium aqueous solution are
37.5mg/mL。
Reference examples 12
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protect liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, hyaluronic acid,
Heparin, Heparan sulfate are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium aqueous solution described in every mL
The hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, institute are dissolved respectively
State streptomysin 20U, the hyaluronic acid 15mg, the heparin 2mg, the Heparan sulfate 2mg, the DMEM/F12 culture
The concentration of base aqueous solution is 37.5mg/mL.
Reference examples 13
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, trehalose, sea
Mosanom is formed with DMEM/F12 culture medium aqueous dissolution, described in DMEM/F12 culture medium aqueous solution dissolves respectively described in every mL
Hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the streptomysin 20U,
The trehalose 2mg, 30 μ g of the sodium alginate, the concentration of the DMEM/F12 culture medium aqueous solution are 37.5mg/mL.
Reference examples 14
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, trehalose, hydroxyl
Propyl cyclodextrin, vitamin E are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium aqueous solution described in every mL
The hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, institute are dissolved respectively
State streptomysin 20U, the trehalose 2mg, 30 μ g of the hydroxypropyl cyclodextrin, 30 μ g of the vitamin E, the DMEM/F12 training
The concentration for supporting base aqueous solution is 37.5mg/mL.
Reference examples 15
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, glucan, hydroxyl
Third methylcellulose is formed with DMEM/F12 culture medium aqueous dissolution, and the difference of DMEM/F12 culture medium aqueous solution described in every mL is molten
Solve the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins 40U, the strepto-
Plain 20U, the bextran 45 mg, the hypromellose 4mg, the concentration of the DMEM/F12 culture medium aqueous solution are
37.5mg/mL。
Reference examples 16
One kind is for fat marrow acquisition suit, unlike the first embodiment: the rouge contained in the protection liquid bottle 2
Fat bone marrow cell protects liquid mainly by hypoxemia protective agent, vitamin C, gentamicin, kanamycins, streptomysin, glucan, sea
Ammonium alginate, lauric monoglyceride are formed with DMEM/F12 culture medium aqueous dissolution, DMEM/F12 culture medium water described in every mL
Solution dissolves the hypoxemia protective agent 8mg, 20 μ g of the vitamin C, the gentamicin 40U, the kanamycins respectively
40U, the streptomysin 20U, the bextran 45 mg, the ammonium alginate 4mg, the lauric monoglyceride 75 μ g, it is described
The concentration of DMEM/F12 culture medium aqueous solution is 37.5mg/mL.
Fat marrow Cell protective solutions safety analysis:
Example 5, embodiment 8, embodiment 11, reference examples 3-6 fat marrow Cell protective solutions, at 40 DEG C of temperature
At ± 2 DEG C, relative humidity is placed 12 months under conditions of being 75% ± 5%, 1 month during test, 3 months, 6 months, 9
The moon, 12 the end of month are separately sampled primary, detect the infection conditions of bacterium, fungi and virus in fat marrow Cell protective solutions, inspection
It tests and the results are shown in Table 1.
1 fat marrow Cell protective solutions safety analysis of table
Wherein: "-" indicates " not having ", " √ " expression " having ".
Comparing embodiment 8 and embodiment 5, reference examples 3-4 fat marrow Cell protective solutions bacterium infection situation it is found that
Ratio gentamicin, kanamycins and streptomysin that parts by weight are 2:2:1 are added in fat marrow Cell protective solutions to be had
Effect improves the ability that fat marrow Cell protective solutions prevent bacterium infection, reduces antibiotic, variation antibiotic or adjustment antibiotic
Weight fraction ratio, can all lead to the reduced capability of fat marrow Cell protective solutions bacterial-infection resisting.
Comparing embodiment 11 and embodiment 8, the fungi of the fat marrow Cell protective solutions of reference examples 5-6 and virus infection shape
Condition is it is found that fat marrow cytoprotection can be effectively prevented in addition benzalkonium chloride and calcium formate in fat marrow Cell protective solutions
Liquid inductance contaminates fungi and virus, reduces one of ingredient, or the one of ingredient of variation, fat marrow Cell protective solutions prevent very
The effect of bacterium and virus infection can weaken.
Impact analysis of the hypoxemia protective agent to cell survival rate under hypoxia condition:
The fat stem cell of identical quantity is respectively put into the fatty bone of embodiment 14, embodiment 17 and reference examples 7-10
Myelocyte is protected in liquid, is placed in the cillin bottle of sterilizing, in 4 DEG C of hypoxemia incubator (1% O2, 5% CO2With 94%
N2) blue dyeing counting fat stem cell survival rate is expected with platform after middle preservation 24 hours, it the results are shown in Table 2.
Cell survival rate under 2 hypoxia condition of table
| Embodiment 14 | Embodiment 17 | Reference examples 7 | Reference examples 8 | Reference examples 9 | Reference examples 10 | |
| Cell survival rate (%) | 85.3 | 94.4 | 51.8 | 70.3 | 87.2 | 85.7 |
From the above results: selecting banaba leaf extract, Herba Oxalidis Corniculatae extract and oxymatrine as hypoxemia
The survival rate of fat stem cell under hypoxia condition can be improved in protective agent；Erythrothioneine and dibutyl hydroxy toluene and great Hua is added
Banaba leaf extract, Herba Oxalidis Corniculatae extract and oxymatrine are used as hypoxemia protective agent together, for improving under hypoxia condition
Fat stem cell survival rate better effect.
Impact analysis of the fat marrow Cell protective solutions to cell survival rate:
Setting test 1-4 and comparative test 1-6, the mesenchymal stem cell of identical quantity is respectively put into different thin
Born of the same parents protect in liquid, are placed in the cillin bottle of sterilizing, save 240 hours at 4 DEG C, during which respectively at 12h, for 24 hours, 48h, 96h,
Mesenchymal stem cell form is observed when 240h, and expects blue dyeing counting mesenchymal stem cell survival rate with platform.
Wherein the fat marrow cell guarantor of embodiment 5, embodiment 8, embodiment 20, embodiment 22 is respectively adopted in test 1-4
Shield liquid makees Cell protective solutions；Comparative test 1 is using physiological saline as Cell protective solutions；Comparative test 2 is trained using DMEM/F12
Base is supported as Cell protective solutions, comparative test 3-6 uses the fat marrow of reference examples 1, reference examples 2, reference examples 11, reference examples 12
Cell protective solutions are shown in Table 3 as Cell protective solutions, test result.
Influence of the 3 variety classes Cell protective solutions of table to cell survival rate
Wherein: A indicates that cell good dispersion degree, uniform in size, form is constant, clear-cut；
B indicates that cell volume becomes larger, differs in size, soft edge, oval or irregular etc. abnormal morphologies occurs.
Above-mentioned test result shows to test 1-4 compared with comparative test 1-4, and mesenchymal stem cell survival rate is bright
It is aobvious higher, and the form of mesenchymal stem cell is also kept as more preferably, it can thus be appreciated that fat marrow provided by the invention is thin
The survival rate that born of the same parents protect liquid that can effectively keep mesenchymal stem cell activity, improve mesenchymal stem cell.
The mesenchymal stem cell survival rate of test 3-4 is apparently higher than test 1-2 and comparative experiments 5-6, illustrates in rouge
Hyaluronic acid, chitin and Heparan sulfate is added in fat bone marrow cell protection liquid to be conducive to improve mesenchymal stem cell
Survival rate reduces one of ingredient, or the one of ingredient of variation, improves the effect meeting of mesenchymal stem cell survival rate
It is deteriorated.
Impact analysis of the fat marrow Cell protective solutions to fat stem cell proliferative capacity
The fat marrow Cell protective solutions that embodiment 8, embodiment 23, reference examples 13 and reference examples 14 are respectively adopted save rouge
Fat stem cell 1h, and by above-mentioned fat stem cell with every cm2The standard for being inoculated with 25 fat stem cells is inoculated on culture plate
It cultivates 7 days afterwards, calculates fat stem cell quantity daily, draw its growth curve, growth curve is as shown in Figure 1.
As can be known from Fig. 1, the proliferation speed for the fat stem cell that the fat marrow Cell protective solutions by embodiment 23 save
Degree is apparently higher than the fat stem cell that the fat marrow Cell protective solutions by embodiment 8, reference examples 13 and reference examples 14 save,
The above results show that trehalose, hydroxypropyl cyclodextrin and sodium alginate are added in fat marrow Cell protective solutions effectively to be mentioned
The growth rate of high-fat stem cell reduces one of ingredient, or the one of ingredient of variation, improves the increasing of fat stem cell
The effect for growing speed can weaken.
Fat marrow Cell protective solutions stability test analysis
Example 8, embodiment 26, reference examples 15-16 fat marrow Cell protective solutions, at 40 DEG C ± 2 DEG C of temperature
Under, relative humidity is placed 12 months under conditions of being 75% ± 5%, 1 month during test, 3 months, 6 months, 9 months, 12
A the end of month is separately sampled primary, detects the character and color of fat marrow Cell protective solutions, and test result is shown in Table 4.
4 fat marrow Cell protective solutions stability test result of table
Wherein: "-" expression does not have, and " √ " is indicated.
The stability of the fat marrow Cell protective solutions of comparing embodiment 26 and embodiment 8 and reference examples 15-16 can
Know, glucan, hypromellose and lauric monoglyceride are added in fat marrow Cell protective solutions effectively to be mentioned
The stability of high-fat bone marrow cell protection liquid, the problems such as avoiding fat marrow Cell protective solutions from generating precipitating, be layered and change colour,
One of ingredient is reduced, or the one of ingredient of variation, the stability of fat marrow Cell protective solutions can weaken.
Finally it should be noted that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although reference
Preferred embodiment describes the invention in detail, those skilled in the art should understand that, it can be to of the invention
Technical solution is modified or replaced equivalently, and without departing from the spirit and scope of the technical solution of the present invention, should all be covered
In scope of the presently claimed invention.
Claims (7)
1. one kind is for fat stem cell and mesenchymal stem cell acquisition suit, it is characterised in that: acquisition set
Dress includes collecting cassette (1), the protection liquid bottle (2) for being provided with fat marrow Cell protective solutions, two collection tubes (3), several ice chests
(4), acquisition suit operation instruction, capture card and a set of bar code, the collecting cassette (1) include box body (101) and the box body
(101) hinged box cover (102) and it is mounted on the collet (103) of box body (101) interior bottom, the collet (103), which is equipped with, to be used
The protection liquid bottle jack (104) of liquid bottle (2) is protected in placement and two collection tube jacks (105) for placing collection tube (3),
The partition (106) around the collet (103), the partition (106) and box body (101) bottom surface are equipped in the box body (101)
Vertically, the ice chest (4) is placed in the cavity formed between the box body (101) side wall and the partition (106), the box
Lid (102) upper surface is additionally provided with the groove (107) for placing acquisition suit operation instruction, capture card and bar code, described recessed
Slot (107) is equipped with and groove (107) matched cover board (108)；The fat marrow cell contained in protection liquid bottle (2) is protected
Protect liquid mainly by hypoxemia protective agent, vitamin C and aminoglycoside antibiotics with DMEM/F12 culture medium aqueous dissolution and
At DMEM/F12 culture medium aqueous solution described in every mL dissolves the hypoxemia protective agent 5-10mg, the vitamin C 10-50 respectively
μ g, the aminoglycoside antibiotics 50-200U, the concentration of the DMEM/F12 culture medium aqueous solution are 30-40mg/mL, institute
State each ingredient that hypoxemia protective agent includes following parts by weight: banaba leaf extract 2-5 Herba Oxalidis Corniculatae extract 2-5 oxidation is bitter
Join alkali 1-2, the aminoglycoside antibiotics is made of gentamicin, kanamycins and streptomysin according to the ratio of 2:2:1.
2. according to claim 1 for fat stem cell and mesenchymal stem cell acquisition suit, feature exists
In: flexible fastening circle (109) are equipped on the protection liquid bottle jack (104) and collection tube jack (105).
3. according to claim 1 for fat stem cell and mesenchymal stem cell acquisition suit, feature exists
In: partition (106) upper end is equipped with inner cover (110), and the inner surface of the inner cover (110) is equipped with and the protection liquid bottle
(2) matched protection liquid bottle cap hole (111) and with the matched collection tube cap bore (112) of the collection tube (3).
4. according to claim 1 for fat stem cell and mesenchymal stem cell acquisition suit, feature exists
In: device for monitoring temperature is additionally provided on the collecting cassette (1).
5. according to claim 1 for fat stem cell and mesenchymal stem cell acquisition suit, feature exists
In: box body (101) bottom is equipped with anti-skid device (113).
6. according to claim 1 for fat stem cell and mesenchymal stem cell acquisition suit, feature exists
In: the junction of the box body (101) and box cover (102) is equipped with fixed device (114).
7. according to claim 1 for fat stem cell and mesenchymal stem cell acquisition suit, feature exists
In: the hypoxemia protective agent further includes each ingredient of following parts by weight:
Erythrothioneine 2-5 dibutyl hydroxy toluene 1-3.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610622783.6A CN106070185B (en) | 2016-08-01 | 2016-08-01 | One kind is for fat marrow acquisition suit |
| PCT/CN2016/094681 WO2018023830A1 (en) | 2016-08-01 | 2016-08-11 | Fatty bone marrow collection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610622783.6A CN106070185B (en) | 2016-08-01 | 2016-08-01 | One kind is for fat marrow acquisition suit |
Publications (2)
| Publication Number | Publication Date |
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| CN109757469A (en) * | 2019-03-20 | 2019-05-17 | 江苏瑞思坦生物科技有限公司 | A kind of adipose tissue transport liquid and preparation method |
| CN109971642A (en) * | 2019-04-30 | 2019-07-05 | 江苏三和生物工程股份有限公司 | A kind of medical instrument for pathogen quick diagnosis |
| CN113230277A (en) * | 2021-05-08 | 2021-08-10 | 陕西鸿瑞康生物科技有限公司 | Stem cell composite preparation and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102433298A (en) * | 2011-11-17 | 2012-05-02 | 安徽农业大学 | Mouse fat stem cell culture solution |
| CN203094812U (en) * | 2012-10-15 | 2013-07-31 | 臻景生物技术（上海）有限公司 | Umbilical cord collection kit |
| CN203698893U (en) * | 2013-12-07 | 2014-07-09 | 黑龙江天晴干细胞有限公司 | Multifunctional adipose collecting and transporting reagent kit |
| CN205120720U (en) * | 2015-06-05 | 2016-03-30 | 军科正源（北京）药物研究有限责任公司 | Kit of anti medicine antibody of short -term test EGFR antibody medicine |
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| US20100075410A1 (en) * | 2008-09-20 | 2010-03-25 | Virendra Desai | Reusable Tissue Biopsy Kit with Padded Cassette |
| CN202603478U (en) * | 2012-05-04 | 2012-12-19 | 臻景生物技术（上海）有限公司 | Kit for preserving adipose tissues during long-term transportation |
| CN203194403U (en) * | 2013-04-21 | 2013-09-18 | 赵静 | Low-temperature living body freezer with temperature display function |
| CN204039387U (en) * | 2014-09-17 | 2014-12-24 | 黑龙江天晴干细胞有限公司 | A kind of acquisition process test kit for obtaining stem cell in tooth |
| CN204625628U (en) * | 2015-02-02 | 2015-09-09 | 中国人民解放军第四五五医院 | Portable biometric sampling box |
| CN206005676U (en) * | 2016-08-01 | 2017-03-15 | 北京世纪劲得生物技术有限公司 | One kind is set with for fat marrow collection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102433298A (en) * | 2011-11-17 | 2012-05-02 | 安徽农业大学 | Mouse fat stem cell culture solution |
| CN203094812U (en) * | 2012-10-15 | 2013-07-31 | 臻景生物技术（上海）有限公司 | Umbilical cord collection kit |
| CN203698893U (en) * | 2013-12-07 | 2014-07-09 | 黑龙江天晴干细胞有限公司 | Multifunctional adipose collecting and transporting reagent kit |
| CN205120720U (en) * | 2015-06-05 | 2016-03-30 | 军科正源（北京）药物研究有限责任公司 | Kit of anti medicine antibody of short -term test EGFR antibody medicine |
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| CN106070185A (en) | 2016-11-09 |
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