CN106167762A - A kind of skin spermatogonium collection suit - Google Patents
A kind of skin spermatogonium collection suit Download PDFInfo
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- CN106167762A CN106167762A CN201610622081.8A CN201610622081A CN106167762A CN 106167762 A CN106167762 A CN 106167762A CN 201610622081 A CN201610622081 A CN 201610622081A CN 106167762 A CN106167762 A CN 106167762A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses the collection suit of a kind of skin spermatogonium, include acquisition component in collecting cassette and box；Collecting cassette includes collecting cassette enclosing cover, collecting cassette inner shell, is located at that the first Magnet inside collecting cassette enclosing cover is corresponding with the first Magnet and fixing pad in being located at the second Magnet outside collecting cassette inner shell and the box be located in collecting cassette inner shell；In box, acquisition component comprises double-level-metal ring, reagent bottle, capture card and ice chest that collection tube mates and the temperature discoloring reagent paper being fixed on double-level-metal ring with collection tube；In box, the ice chest draw-in groove of pad one end fixed by fixing pad in being provided with collection tube draw-in groove, double-level-metal ring draw-in groove, reagent bottle draw-in groove, capture card draw-in groove and being located at box.Equipped with Cell protective solutions in reagent bottle, aminoglycoside antibiotics DMEM culture medium aqueous dissolution is mainly formed by protection liquid.This collection is easy to use with suit, and protection liquid fully simulates internal milieu, it is possible to significantly improve the survival rate of cell.
Description
Technical field
The present invention relates to a kind of cell collection suit, be specifically related to a kind of skin spermatogonium collection suit.
Background technology
In the clinic of current China and cell research experiment, the collection of cell, long-term preservation and use are that space-time separates,
After the cells ex vivo gathered the harshest to the requirement of external environment condition, particularly cell collection and long-term preserve between cell turn
During fortune, due to cell have not gone through long-term preservation process the bad cell viability that can reduce of external environment condition, determine gathers successfully and
No.Transport variation of ambient temperature in transportation after gathering to facilitate, monitor accurately, it is not known that transportation has simultaneously
Do not cause cell inactivation because ambient temperature is defective.Therefore, cell collection, the environment created are carried out easily and efficiently
Cell is transported through, and it is monitored, be a critically important problem in the clinic of China and cell research experiment.
Summary of the invention
The invention provides one and gather suit, cell collection, and the environment pair created can be carried out easily and efficiently
Cell is transported through, and gathers the protection liquid in suit, it is further provided the suitable and stable Conservation environment of skin spermatogonium,
The cell viability after skin spermatogonium preserves can be significantly improved.Concrete technical scheme is as follows:
A kind of skin spermatogonium collection suit, includes acquisition component in collecting cassette and box；
Described collecting cassette includes collecting cassette enclosing cover, collecting cassette inner shell, is located at the first Magnet inside collecting cassette enclosing cover and
One Magnet is corresponding and is located at fixing pad in the second Magnet outside collecting cassette inner shell and the box be located in collecting cassette inner shell；
In described box, acquisition component comprises double-level-metal ring, reagent bottle, capture card and the ice that collection tube mates with collection tube
Box, the temperature discoloring reagent paper being fixed on double-level-metal ring；
In described box fixing pad be provided with collection tube draw-in groove, double-level-metal ring draw-in groove, reagent bottle draw-in groove, capture card draw-in groove and
The ice chest draw-in groove of fixing pad one end in being located at box.
Preferably, described collecting cassette enclosing cover and collecting cassette inner shell material are PC plastics.
Preferably, described collection tube is PC test tube with a scale and pipe cap.
Preferably, described double-level-metal ring material is aluminum.
Preferably, in described box acquisition component also to include bar code a set of.
Preferably, in described box, acquisition component also includes collection suit operation instruction portion.
Preferably, equipped with Cell protective solutions in described reagent bottle, described protection liquid is mainly aminoglycoside antibiotics
Form with DMEM culture medium aqueous dissolution, aminoglycoside antibiotics described in every milliliter of described DMEM culture medium aqueous dissolution
50-200 unit, in described DMEM culture medium aqueous solution, the concentration of DMEM culture medium is 10-25g/L, and described aminoglycoside resists
One or more in gentamycin, kanamycin, amikacin or tobramycin of raw element.
Preferably, described protection liquid also includes HbO2 Oxyhemoglobin, every milliliter of described DMEM culture medium aqueous dissolution institute
State HbO2 Oxyhemoglobin 400-1000 μ g.
Preferably, described protection liquid also includes that one or more in vitamin E or glutathion, every milliliter of DMEM are cultivated
Base aqueous dissolution vitamin E 0.25 μm ol and glutathion 0.25 μm ol.
Preferably, described protection liquid also includes Tween 80, palmitamide, potassium alginate and Hamposyl L, every milliliter
Described DMEM culture medium aqueous solution dissolves Tween 80 0.5 μ L, palmitamide 60 μ g, potassium alginate 30 μ g, lauroyl flesh ammonia respectively
Acid 150 μ g.
Preferably, described protection liquid also includes citric acid, acetone acid, Fumaric acid and S-acetyl-coenzyme-A, described in every milliliter
DMEM culture medium aqueous solution dissolves citric acid 1-10 μ g, acetone acid 50-80 μ g, Fumaric acid 10-20 μ g and acetylcoenzyme respectively
A80-150μg。
The invention has the beneficial effects as follows: the collection suit that the present invention provides is easy to use, it is simple to Skin Cell or essence are former
Protection liquid in the storage of cell and transport, and reagent bottle fully simulates internal milieu, is more suitable for Skin Cell and essence is former
The existence of cell, it is possible to significantly improve the survival rate of above cell.
Accompanying drawing explanation
Fig. 1 is the upward view of embodiment 1 collecting cassette enclosing cover；
Fig. 2 is the collection suit top view that embodiment 1 opens collecting cassette enclosing cover；In figure, 1 collecting cassette enclosing cover, 2 collecting cassettes
Fixing pad, 11 collection tube draw-in grooves, 12 double-level-metal ring draw-in grooves, 13 reagent bottle cards in inner shell, 3 first Magnet, 4 second Magnet, 5 boxes
Groove, 14 capture card draw-in grooves, 15 ice chest draw-in grooves.
Detailed description of the invention
Embodiment 1
A kind of skin spermatogonium collection suit, includes acquisition component in collecting cassette and box；
Described collecting cassette include collecting cassette enclosing cover 1, collecting cassette inner shell 2, be located at the first Magnet 3 inside collecting cassette enclosing cover 1,
Corresponding with the first Magnet 3 and fix pad 5 in being located at the second Magnet 4 outside collecting cassette inner shell and box；
In described box, acquisition component comprises double-level-metal ring that collection tube mates, reagent bottle, capture card, ice with collection tube
Box and the temperature discoloring reagent paper being fixed on double-level-metal ring；
In described box, fixing pad 5 is provided with collection tube draw-in groove 11, double-level-metal ring draw-in groove 12, reagent bottle draw-in groove 13, gathers
Card draw-in groove 14 and the ice chest draw-in groove 15 of fixing pad 5 one end in being located at box.
Gather the collection suit collecting cassette enclosing cover of front opening skin spermatogonium, take ready ice chest and put into ice chest card
Groove.
Then take out collection tube, then the skin spermatogonium of collection is put in collection tube, often pipe 25ml.By in reagent bottle
Protection liquid pour in collection tube in the ratio of 1:1, tighten collection lid.Then on collection tube, labelling is carried out with marking pen.
Then the temperature discoloring reagent paper being fixed in double-level-metal ring mated with collection tube is checked, after confirming normally, will
The double-level-metal ring mated with collection tube being fixed with temperature discoloring reagent paper is contained on collection tube, and becket is close to collection tube.
When collection tube needs temporarily to preserve, put into 4 DEG C of refrigerators, ensure that collection tube lid is tightened, collection tube is upright, surely simultaneously
Fixed, can preserve within 12 hours.
Then on capture card, fill in donor data and collection information in detail.Before gathering suit handing-over transport, replace and gather
Set ice chest draw-in groove in ice chest, by collection tube, fill in complete capture card by origin-location put into collection suit in, cover collection
Box enclosing cover, it is ensured that be located at the first Magnet inside collecting cassette enclosing cover and second magnetic be located at inner shell outside corresponding with the first Magnet
Ferrum attracts each other.
Transport completes, and opens the collection suit of skin spermatogonium, checks that mate with collection tube is fixed on becket
Temperature discoloring reagent paper, confirm normal after, complete handing-over, measure carry out after cell viability meets the requirements follow-up long-term preserve or
Person breeds.If variable color reagent paper displays temperature is higher than the requirement temperature to environment, measure cell viability undesirable then for gathering
Failure, needs the skin spermatogonium abandoning gathering.
The collection suit of the skin spermatogonium of the present embodiment, easy to use, cheap.It is located in collecting cassette enclosing cover
First Magnet and be located at second Magnet inner shell outside corresponding with the first Magnet attract each other, and reduce in transportation of knowing clearly and adopt
Collection box enclosing cover is because rocking or other probability that is unexpected and that be opened.Ice in ice chest provides the low temperature environment in collecting cassette, ice chest
Draw-in groove is positioned at one end rather than the bottom of the fixing pad of box, if haulage time is longer, in the case of ice is inadequate, can not take
Carry out the replacement of ice chest in going out box in the case of fixing pad, can preferably ensure low temperature environment.Temperature discoloring reagent paper monitoring fortune
Variations in temperature during defeated.Protect temperature discoloring reagent paper to be easily received water or the pollution of other impurity and lost efficacy, be not suitable for directly
It is attached to monitoring problem on collection tube.Common plastic protective layer with insulation blocking temperature discoloring reagent paper, but also can completely cut off simultaneously
Heat, heat conductivity is bad, and the temperature of temperature discoloring reagent paper and the temperature of collection tube have a certain distance.The bilayer of the present embodiment
Becket sandwich protection temperature discoloring reagent paper, isolation make its not by water or other pollute, be close to collection tube simultaneously, preferably transmit
Heat so that temperature discoloring reagent paper temperature is closer to the temperature with collection tube.
The collection suit internal temperature measure of the change of skin spermatogonium, first takes the ice chest prepared, puts into ice chest draw-in groove,
Put into thermometer, after pre-cooling 30 minutes, observe the temperature of thermometer through transparent collecting cassette and record every 30 minutes records one
Secondary temperature.Gather suit internal temperature in 12 hours and be consistently higher than 4 degrees Celsius.
Gathering 12 hours Dynamic Curves of suit inner skin spermatogonium to measure, mtt assay measures cell viability (" Wang Zhirong, height
Fine jade, Ma Chuanxin, Li Yan girl's .MTT method mensuration escherichia coli viable count experimentation, ACTA Scientiae Circumstantiae, 2011,31 (12):
2642-1650.), sampling in every 30 minutes once, with the vigor of for the first time sampling for 100%.
Embodiment 2
The present embodiment is with the difference of embodiment 1, and in described box, also to include bar code a set of for acquisition component.
During use, directly bar code is attached on collection tube, so can time-consuming, add fast speed, it is thus also avoided that note
Number risk that a labelling is wiped in transit by mistake.
Embodiment 3
The present embodiment is with the difference of embodiment 1, and in described box, acquisition component also includes collection suit operation instruction
A.
For not using this skin spermatogonium to gather the people of suit in the past, can carry out ripe according to operation instruction
The operation practiced.
Embodiment 4
The present embodiment is with the difference of embodiment 1, and described collection tube is PC test tube with a scale and pipe cap, described double
Layer becket material is aluminum, equipped with Cell protective solutions in described reagent bottle.
Embodiment 5 protects the preparation of liquid, use and cell viability to measure
Protection liquid 1
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly molten for gentamycin DMEM culture medium aqueous solution
Solution forms, every milliliter of described DMEM culture medium aqueous dissolution gentamycin 100 unit, described DMEM culture medium aqueous solution dense
Degree is 16.9g/L.
Protection liquid 2
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly molten for kanamycin DMEM culture medium aqueous solution
Solution forms, every milliliter of described DMEM culture medium aqueous dissolution kanamycin 100 unit, described DMEM culture medium aqueous solution dense
Degree is 16.9g/L.
Protection liquid 3
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly molten for amikacin DMEM culture medium aqueous solution
Solution forms, every milliliter of described DMEM culture medium aqueous dissolution amikacin 100 unit, described DMEM culture medium aqueous solution dense
Degree is 16.9g/L.
Protection liquid 4
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, kanamycin and tobramycin
Form with DMEM culture medium aqueous dissolution, every milliliter of described DMEM culture medium aqueous solution dissolve respectively gentamycin 100 unit,
Kanamycin 20 unit, tobramycin 20 unit, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 5
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly molten for tobramycin DMEM culture medium aqueous solution
Solution forms, every milliliter of described DMEM culture medium aqueous dissolution tobramycin 100 unit, described DMEM culture medium aqueous solution dense
Degree is 10.0g/L.
Protection liquid 6
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly molten for gentamycin DMEM culture medium aqueous solution
Solution forms, every milliliter of described DMEM culture medium aqueous dissolution gentamycin 100 unit, described DMEM culture medium aqueous solution dense
Degree is 25.0g/L.
Protection liquid 7
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin and HbO2 Oxyhemoglobin DMEM
Culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, oxygenated blood respectively
Lactoferrin 400 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 8
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin and HbO2 Oxyhemoglobin DMEM
Culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, oxygenated blood respectively
Lactoferrin 600 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 9
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin and HbO2 Oxyhemoglobin DMEM
Culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, oxygenated blood respectively
Lactoferrin 1000 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 10
A kind of skin stem spermatogonium protection liquid, gentamycin and vitamin E DMEM are mainly cultivated by this protection liquid
Base aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, vitamin E 0.1 respectively
μm ol, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 11
A kind of skin stem spermatogonium protection liquid, gentamycin, vitamin E and glutathion are mainly used by this protection liquid
DMEM culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, dimension respectively
Raw element E0.3 μm ol, glutathion 0.3 μm ol, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 12
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, HbO2 Oxyhemoglobin, vitamin
E and glutathion DMEM culture medium aqueous dissolution form, and every milliliter of described DMEM culture medium aqueous solution dissolves celebrating greatly respectively
Mycin 100 unit, HbO2 Oxyhemoglobin 600 μ g, vitamin E 0.25 μm ol, glutathion 0.25 μm ol, described DMEM culture medium
The concentration of aqueous solution is 16.9g/L.
Protection liquid 13
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, HbO2 Oxyhemoglobin, vitamin
C and vitamin E DMEM culture medium aqueous dissolution form, and it is the most mould that every milliliter of described DMEM culture medium aqueous solution dissolves celebrating respectively
Element 100 units, HbO2 Oxyhemoglobin 600 μ g, vitamin C 0.5 μm ol, vitamin E 0.5 μm ol, described DMEM culture medium is water-soluble
The concentration of liquid is 16.9g/L.
Protection liquid 14
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, Tween 80, palmitamide, sea
Potassium alginate and Hamposyl L DMEM culture medium aqueous dissolution form, and every milliliter of described DMEM culture medium aqueous solution is respectively
Dissolve gentamycin 100 unit, Tween 80 0.5 μ g, palmitamide 80 μ g, potassium alginate 50 μ g, Hamposyl L 100 μ g,
The concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 15
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, HbO2 Oxyhemoglobin, tween
80, palmitamide, potassium alginate and Hamposyl L DMEM culture medium aqueous dissolution form, every milliliter of described DMEM training
Support base aqueous solution and dissolve gentamycin 100 unit, HbO2 Oxyhemoglobin 600 μ g, Tween 80 1.0 μ g, palmitamide 20 μ respectively
G, potassium alginate 10 μ g, Hamposyl L 200 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 16
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, HbO2 Oxyhemoglobin, vitamin
E, glutathion, Tween 80, palmitamide, potassium alginate and Hamposyl L DMEM culture medium aqueous dissolution form,
Every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, HbO2 Oxyhemoglobin 600mg, vitamin respectively
E0.25 μm ol, glutathion 0.25 μm ol, Tween 80 0.5 μ g, palmitamide 60 μ g, potassium alginate 30 μ g, lauroyl flesh ammonia
Acid 150 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 17
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, citric acid, acetone acid, Rhizoma Corydalis
Rope acid and S-acetyl-coenzyme-A DMEM culture medium aqueous dissolution form, and every milliliter of described DMEM culture medium aqueous dissolution is respectively
Gentamycin 100 unit, citric acid 10 μ g, acetone acid 80 μ g, Fumaric acid 10 μ g and S-acetyl-coenzyme-A 80 μ g, described DMEM trains
The concentration supporting base aqueous solution is 16.9g/L.
Protection liquid 18
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, HbO2 Oxyhemoglobin, vitamin
E, glutathion, citric acid, acetone acid, Fumaric acid and S-acetyl-coenzyme-A DMEM culture medium aqueous dissolution form, every milli
Rise described DMEM culture medium aqueous solution and dissolve gentamycin 100 unit, HbO2 Oxyhemoglobin 600 μ g, vitamin E 0.25 μ respectively
Mol, glutathion 0.25 μm ol, citric acid 1 μ g, acetone acid 50 μ g, Fumaric acid 20 μ g and S-acetyl-coenzyme-A 150 μ g, described
The concentration of DMEM culture medium aqueous solution is 16.9g/L.
Protection liquid 19
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, HbO2 Oxyhemoglobin, vitamin
E, glutathion, Tween 80, palmitamide, potassium alginate, Hamposyl L, citric acid, acetone acid, Fumaric acid and acetyl
Coenzyme A DMEM culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 respectively
Unit, HbO2 Oxyhemoglobin 600 μ g, vitamin E 0.25 μm ol, glutathion 0.25 μm ol, Tween 80 0.5 μ g, palmitamide
60 μ g, potassium alginate 30 μ g, Hamposyl L 150 μ g, citric acid 4 μ g, acetone acid 60 μ g, Fumaric acid 15 μ g and acetyl are auxiliary
Enzyme A 120 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Comparison liquid 1
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly penicillin DMEM culture medium aqueous dissolution
Forming, every milliliter of described DMEM culture medium aqueous dissolution penicillin 100 unit, the concentration of described DMEM culture medium aqueous solution is
16.9g/L。
Comparison liquid 2
A kind of skin stem spermatogonium protection liquid, gentamycin and transferrins DMEM are mainly cultivated by this protection liquid
Base aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, transferrins respectively
400 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Comparison liquid 3
A kind of skin stem spermatogonium protection liquid, gentamycin, vitamin E and glutathion are mainly used by this protection liquid
DMEM culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 unit, dimension respectively
Raw element E0.1 μm ol, glutathion 0.9 μm ol, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Comparison liquid 4
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, sorbester p17, palmitamide, sea
Potassium alginate and Hamposyl L DMEM culture medium aqueous dissolution form, and every milliliter of described DMEM culture medium aqueous solution is respectively
Dissolve gentamycin 100 unit, sorbester p17 0.5 μ g, palmitamide 80 μ g, potassium citrate 50 μ g, Hamposyl L 100 μ g,
The concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Comparison liquid 5
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, Tween 80, palmitamide and sea
Potassium alginate DMEM culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin respectively
100 units, Tween 80 0.5 μ g, palmitamide 80 μ g, potassium alginate 50 μ g, the concentration of described DMEM culture medium aqueous solution is
16.9g/L。
Comparison liquid 6
A kind of skin stem spermatogonium protection liquid, this protection liquid mainly by gentamycin, potassium citrate, Sodium Pyruvate,
Fumaric acid and S-acetyl-coenzyme-A DMEM culture medium aqueous dissolution form, and every milliliter of described DMEM culture medium aqueous solution is respectively
Dissolve gentamycin 100 unit, potassium citrate 10 μ g, Sodium Pyruvate 80 μ g, Fumaric acid 10 μ g and S-acetyl-coenzyme-A 80 μ g, institute
The concentration stating DMEM culture medium aqueous solution is 16.9g/L.
Comparison liquid 7
A kind of skin stem spermatogonium protection liquid, this protection liquid is mainly gentamycin, citric acid, acetone acid and Rhizoma Corydalis
Rope acid DMEM culture medium aqueous dissolution forms, and every milliliter of described DMEM culture medium aqueous solution dissolves gentamycin 100 respectively
Unit, citric acid 10 μ g, acetone acid 80 μ g and Fumaric acid 10 μ g, the concentration of described DMEM culture medium aqueous solution is 16.9g/L.
Test example 1
Take the sterilizing protection liquid being stored in 4 DEG C of refrigerators, press 1:1 volume ratio with Skin Cell or the spermatogonium just gathered
Mix homogeneously, is placed in 4 DEG C of environment, and sampling should be carried out for different time points, measures cell viability with mtt assay, with sampling for the first time
Vigor be 100%.
The test of this group is main investigates aminoglycoside antibiotics and the impact of DMEM concentration, to compare what liquid 1 preserved respectively
Skin Cell and spermatogonium are comparison 1 group, to protect Skin Cell and spermatogonium that liquid 2-6 preserves respectively for experiment 1-5
Group, each experimental group and the Skin Cell of matched group and spermatogonium enter test procedure the most after 24h, adjust Skin Cell and
The density of spermatogonium is 1 × 106cells/mL.By cell suspension: 0.4% trypan blue=3:1 (v:v) fully mixes, and takes
20 μ L cell suspension add in cell counting count board, detect each experimental group and matched group protection liquid with Countstar cell counter
Endepidermis stem cell and the motility rate of stem spermatogonium, result is such as
Shown in table 1.
Table 1 test example 1 respectively protects liquid and the Cell viability result compareing liquid
As shown in Table 1, protection liquid 1-6 preserves respectively Skin Cell and the Cell viability ratio of spermatogonium use comparison
Skin Cell and the Cell viability of spermatogonium that the protection liquid that liquid 1 group uses preserves respectively are high；The protection liquid that protection liquid 4 uses
The Skin Cell and the Cell viability of spermatogonium that preserve respectively are protected respectively than the protection liquid using protection liquid 1-3,5-6 to use
The Skin Cell deposited and the Cell viability of spermatogonium are high.
It follows that the aminoglycoside antibiotics in liquid of protecting that the present invention provides has changed the antibiotic such as penicillin into
After, the motility rate of Skin Cell or spermatogonium significantly reduces, and aminoglycoside antibiotics uses gentamycin, kanamycin
After the mixture of tobramycin, the Skin Cell of preservation and the Cell viability of spermatogonium are higher.
Test example 2
The main impact investigating HbO2 Oxyhemoglobin of this group test, former to compare Skin Cell that liquid 2 preserves respectively and essence
Cell is comparison 2 groups, to protect Skin Cell and spermatogonium that liquid 1,7 and 8 preserves respectively for experiment 7-9 group, detects process
Such as test example 1, preserve the testing result such as table 2 after 24h.
Table 2 test example 2 respectively protects liquid and the Cell viability result compareing liquid
As shown in Table 2, protection liquid 7,8 preserves respectively Skin Cell and the Cell viability ratio of spermatogonium use comparison
Skin Cell and the Cell viability of spermatogonium that the protection liquid that liquid 2 groups uses preserves respectively are high；The skin that protection liquid 8 preserves respectively
The Cell viability of skin cell and spermatogonium is lived than the cell of the Skin Cell using protection liquid 7 to preserve respectively and spermatogonium
Rate is high.
It follows that after the HbO2 Oxyhemoglobin in the protection liquid of present invention offer has changed transferrins into, Skin Cell
Or the motility rate of spermatogonium significantly reduces, and when the concentration of HbO2 Oxyhemoglobin is 600 μ g/mL, the skin that protection liquid preserves respectively
The Cell viability Gao Genggao of skin cell and spermatogonium.
Test example 3
The main impact investigating antioxidant of this group test, to compare Skin Cell and the spermatogonium that liquid 3 preserves respectively
For matched group, protection liquid 7,10-12 are experimental group, detect process such as test example 1, preserve the testing result after 48h
Such as table 2.
Table 3 test example 3 respectively protects liquid and the Cell viability result compareing liquid
As shown in Table 3, protection liquid 10-12 preserves respectively Skin Cell and the Cell viability of spermatogonium are protected than using
The Cell viability protecting liquid 7, the Skin Cell protecting liquid to preserve respectively of comparison liquid 3 groups use and spermatogonium is high；
Skin Cell and the Cell viability ratio of spermatogonium that protection liquid 12 preserves respectively use protection liquid 10,11 respectively
The Skin Cell preserved and the Cell viability of spermatogonium are high.
It follows that the antioxidant in liquid of protecting that the present invention provides can significantly improve Skin Cell or spermatogonium
Motility rate.
Test example 4
The Skin Cell preserved respectively with comparison liquid 4-7 and spermatogonium, as matched group, protect liquid 4, protection liquid 7, protection
Liquid 10, protection liquid 14,16, protection liquid 17 and protection liquid 19 are experimental group, and each experimental group and the Skin Cell of matched group and essence are former
Cell enters test procedure respectively after 24h, 48h, 96h, 240h, 480h, and the density adjusting Skin Cell and spermatogonium is equal
It is 1 × 106cells/mL.By cell suspension: 0.4% trypan blue=3:1 (v:v) fully mixes, take 20 μ L cell suspension and add
In cell counting count board, detect each experimental group and matched group protection liquid endepidermis stem cell and essence with Countstar cell counter
The motility rate of former stem cell, result is as shown in table 4.
Table 4 test example 4 respectively protects liquid and the Cell viability result compareing liquid
As shown in Table 4, protection liquid 14,16-17 and protection liquid 19 preserve respectively Skin Cell and the cell of spermatogonium
Motility rate is than use protection liquid 1, protection liquid 7 and protection liquid 10 to compare Skin Cell that liquid 4-7 preserves respectively and spermatogonium
Cell viability is high；Skin Cell and the time ratio of spermatogonium that protection liquid 17 and protection liquid 19 preserve respectively use protection liquid
The holding time of 14 and 16 is long.
It follows that Tween 80, palmitamide, potassium alginate and the Hamposyl L in the protection liquid of present invention offer
Compositions can significantly improve the motility rate of Skin Cell or spermatogonium, citric acid, acetone acid, Fumaric acid and S-acetyl-coenzyme-A
Compositions can significantly improve protection liquid to Skin Cell or the holding time of spermatogonium.
Claims (10)
1. the collection suit of a skin spermatogonium, it is characterised in that: include acquisition component in collecting cassette and box；
Described collecting cassette includes collecting cassette enclosing cover (1), collecting cassette inner shell (2), is located at the first Magnet inside collecting cassette enclosing cover (1)
(3) corresponding with the first Magnet (3) and be located at collecting cassette inner shell (2) outside the second Magnet (4) and be located at collecting cassette inner shell (2)
Fixing pad (5) in interior box；
In described box acquisition component comprise double-level-metal ring that collection tube mates with collection tube, reagent bottle, capture card, ice chest and
The temperature discoloring reagent paper being fixed on double-level-metal ring；
In described box fixing pad (5) be provided with collection tube draw-in groove (11), double-level-metal ring draw-in groove (12), reagent bottle draw-in groove (13),
Capture card draw-in groove (14) and the ice chest draw-in groove (15) of fixing pad (5) one end in being located at box.
2. the collection suit of skin spermatogonium as claimed in claim 1, it is characterised in that described collecting cassette enclosing cover (1)
It is PC plastics with collecting cassette inner shell (2) material.
3. the collection suit of skin spermatogonium as claimed in claim 1, it is characterised in that described collection tube is with a scale
PC test tube with pipe cap.
4. the collection suit of skin spermatogonium as claimed in claim 1, it is characterised in that described double-level-metal ring material is
Aluminum.
5. the collection suit of skin spermatogonium as claimed in claim 1, it is characterised in that in described box, acquisition component is also wrapped
A set of containing bar code.
6. the collection suit of skin spermatogonium as claimed in claim 1, it is characterised in that in described box, acquisition component is also wrapped
Containing gathering, suit operation instruction is a.
7. the collection suit of skin spermatogonium as claimed in claim 1, it is characterised in that equipped with cell in described reagent bottle
Protection liquid, aminoglycoside antibiotics DMEM culture medium aqueous dissolution is mainly formed by described protection liquid, every milliliter of institute
State aminoglycoside antibiotics 50-200 unit described in DMEM culture medium aqueous dissolution, in described DMEM culture medium aqueous solution
The concentration of DMEM culture medium is 10-25g/L, and described aminoglycoside antibiotics is selected from gentamycin, kanamycin, amikacin
Or one or more in tobramycin.
8. the collection suit of skin spermatogonium as claimed in claim 7, it is characterised in that described protection liquid also includes oxygenate
Hemoglobin, HbO2 Oxyhemoglobin 400-1000 μ g described in every milliliter of described DMEM culture medium aqueous dissolution.
9. the collection suit of skin spermatogonium as claimed in claim 7, it is characterised in that described protection liquid also includes that dimension is raw
One or more in element E or glutathion, every milliliter of DMEM culture medium aqueous dissolution vitamin E 0.25 μm ol and gluathione
Peptide 0.25 μm ol.
10. the collection suit of skin spermatogonium as claimed in claim 7, it is characterised in that described protection liquid also includes telling
Temperature 80, palmitamide, potassium alginate and Hamposyl L, every milliliter of described DMEM culture medium aqueous solution dissolves Tween 80 respectively
0.5 μ L, palmitamide 60 μ g, potassium alginate 30 μ g, Hamposyl L 150 μ g.
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| CN202015626U (en) * | 2011-03-15 | 2011-10-26 | 高涵 | Portable kit for high school chemistry experiments |
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| CN203283599U (en) * | 2012-12-05 | 2013-11-13 | 江苏和泽生物科技有限公司 | Placenta collection set |
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| CN105062959A (en) * | 2015-09-20 | 2015-11-18 | 领航干细胞再生医学工程有限公司 | Isolated culture method of human amnia mesenchymal stem cells |
| CN105532641A (en) * | 2015-12-28 | 2016-05-04 | 贵州泰邦生物制品有限公司 | Placenta preserving fluid and use method thereof |
| CN206014907U (en) * | 2016-08-01 | 2017-03-15 | 北京世纪劲得生物技术有限公司 | A kind of collection suit of skin spermatogonium |
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2016
- 2016-08-01 CN CN201610622081.8A patent/CN106167762B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202015626U (en) * | 2011-03-15 | 2011-10-26 | 高涵 | Portable kit for high school chemistry experiments |
| CN202738683U (en) * | 2012-07-25 | 2013-02-20 | 臻景生物技术（上海）有限公司 | Tissue cryopreservation liquid kit |
| CN203283599U (en) * | 2012-12-05 | 2013-11-13 | 江苏和泽生物科技有限公司 | Placenta collection set |
| 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 |
| CN105062959A (en) * | 2015-09-20 | 2015-11-18 | 领航干细胞再生医学工程有限公司 | Isolated culture method of human amnia mesenchymal stem cells |
| CN105532641A (en) * | 2015-12-28 | 2016-05-04 | 贵州泰邦生物制品有限公司 | Placenta preserving fluid and use method thereof |
| CN206014907U (en) * | 2016-08-01 | 2017-03-15 | 北京世纪劲得生物技术有限公司 | A kind of collection suit of skin spermatogonium |
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| CN106167762B (en) | 2018-06-22 |
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