CN107306939A - A kind of cells frozen storing liquid for BMDC - Google Patents
A kind of cells frozen storing liquid for BMDC Download PDFInfo
- Publication number
- CN107306939A CN107306939A CN201710635539.8A CN201710635539A CN107306939A CN 107306939 A CN107306939 A CN 107306939A CN 201710635539 A CN201710635539 A CN 201710635539A CN 107306939 A CN107306939 A CN 107306939A
- Authority
- CN
- China
- Prior art keywords
- dcs
- agent
- storing liquid
- frozen storing
- cells frozen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Biophysics (AREA)
- Physiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of cells frozen storing liquid for BMDC; freezing protective agent, cytotostatic agent and the anti-settling agent that effective dose is added in RPMI 1640 culture mediums are uniformly mixed; freezing protective agent is dimethyl sulfoxide (DMSO), and cytotostatic agent is people's gamma Globulin；Anti-settling agent is many PLLs, and the molecular weight ranges of many PLLs are 37 ten thousand；The concentration expressed in percentage by volume of dimethyl sulfoxide (DMSO) is 1 3%.Cells frozen storing liquid of the present invention is used to that when freezing DCs DCs cryopreservation resuscitation rate and immunocompetence can be ensured, freeze cryopreservation resuscitation rate and immunocompetence in 1 year reduces without obvious；People's gamma Globulin is cytotostatic agent, can maintain DCs immunocompetence；Many PLLs are anti-settling agent, it is to avoid storage process people gamma Globulin is settled to frozen stock solution bottom, meanwhile, many PLLs have the effect of part freezing protective agent, additionally assist in maintaining DCs immunocompetence.
Description
Technical field
The invention belongs to field of cell culture, and in particular to a kind of cells frozen storing liquid for BMDC.
Background technology
It is clinical often to need a large amount of ripe BMDCs (DCs), but natural DCs contents are few in body, it is difficult to meet clinical
The need for, can only largely it be obtained by vitro culture.Mature DCs need large amount of cell factor to maintain, and spend high, easy dirt
Dye, is unfavorable for DCs clinical practice, thus can by the DCs freezen protectives prepared, it is necessary to when recover.
In order to keep freezing DCs anabiosis rate and immunocompetence, select suitable freezing and storing method of crucial importance.
At present, DCs cryopreservation methods are using containing 10% dimethyl sulfoxide (DMSO) (DMSO), 20% hyclone (FCS)
The step of frozen stock solution two cools.This method is clearly disadvantageous in the presence of two：
First, FCS complicated component, pollutant can be introduced into DCs and original of causing a disease, and security is difficult control；At present have into
Mouth frozen stock solution is free of FCS (the Cellbanker cells frozen storing liquids produced such as ZENOAQ companies of Japan), but expensive, applies
Cost is too high, limits its use in clinic, especially developing country and undeveloped country；
Second, although DMSO is conventional freezing protective agent, but this does not represent it will not to cause to freeze damage to cell (deep
DMSO cytotoxicity is suppressed during low temperature, and action is fast during recovery, washes dimethyl sulfoxide (DMSO) off as early as possible, otherwise can cause to thin
The serious poison of born of the same parents；But, one skilled in the art will appreciate that speed is fast again, cell toxicant is inevitable), this damage limits multiple
The further raising of Soviet Union's rate；Unless the new reagent that can substitute DMSO of exploitation or reduction DMSO ratio.
In order to reduce the DCs monopolization for freezing cost, breaking import frozen stock solution, applicant is directed to exploitation and possessed from main product
The DCs frozen stock solutions of power, cost is reduced on the premise of DCs cryopreservation resuscitations rate and immunocompetence is ensured.
The content of the invention
It is contemplated that reduction DCs freezes cost, breaking the monopolization of import frozen stock solution, there is provided a kind of economic, practical, height
The DCs frozen stock solutions of effect, cost is reduced on the premise of DCs cryopreservation resuscitations rate and immunocompetence is ensured.
The present invention is achieved by following technical solution：
A kind of cells frozen storing liquid based on RPMI-1640 nutrient solutions, adds the jelly of effective dose in RPMI-1640 nutrient solutions
Deposit protective agent, cytotostatic agent and anti-settling agent to be uniformly mixed, freezing protective agent is dimethyl sulfoxide (DMSO), the cytotostatic agent
For people's gamma Globulin；The anti-settling agent is poly-l-lysine, and the molecular weight ranges of poly-l-lysine are 3-7 ten thousand；It is described
The concentration expressed in percentage by volume of dimethyl sulfoxide (DMSO) is 1-3%.
Preferably, the mass concentration of people's gamma Globulin is 8-12 μ g/mL.
Preferably, mass concentration of the poly-l-lysine in cells frozen storing liquid is 20-30 μ g/mL.
Preferably, antibiotic is also contained in the cells frozen storing liquid.
Preferably, antibiotic is gentamicin, and concentration is 50U/mL.
Above-mentioned cells frozen storing liquid freezes the application of aspect in BMDC.
Advantage of the present invention：
The cells frozen storing liquid that the present invention is provided is used to that when freezing DCs DCs cryopreservation resuscitation rate and immunocompetence can be ensured,
Freeze cryopreservation resuscitation rate and immunocompetence in 1 year reduces without obvious；In cells frozen storing liquid of the present invention, dimethyl sulfoxide (DMSO) is to freeze guarantor
Agent is protected, the damage that low temperature is caused to DCs is reduced；People's gamma Globulin is cytotostatic agent, can maintain DCs immunocompetence；It is many
Poly-L-Lysine is anti-settling agent, it is to avoid storage process people gamma Globulin is settled to frozen stock solution bottom, meanwhile, poly-l-lysine
Effect with part freezing protective agent, the presence of poly-l-lysine causes DMSO volumetric concentration from the 10- of prior art
20% can be reduced to the 1-3% of the present invention and still be avoided that the damage that low temperature is caused to DCs；In addition, poly-l-lysine
Additionally assist in maintaining DCs immunocompetence.The cells frozen storing liquid that the present invention is provided freezes effect and ZENOAQ companies of Japan to DCs
The Cellbanker cells frozen storing liquids of production are basically identical, but cost only has about the 1/10 of the import frozen stock solution.
Brief description of the drawings
Fig. 1 is cryopreservation resuscitation rates (%) of the DCs behind 3,6,9 and December；
Fig. 2 is killing rates (%) of the DC-CIK to K562.
Embodiment
In order to preferably explain technical scheme, it is further described with reference to specific embodiment.In embodiment
The experiment material do not emphasized especially is normal experiment material, belongs to the category that those skilled in the art are easily obtained.
Embodiment 1：The preparation of cells frozen storing liquid
Reagent source：
RPMI-1640 nutrient solutions, Gibco；
Dimethyl sulfoxide (DMSO), lark prestige Science and Technology Ltd.；
People's gamma Globulin (No. CAS：9007-83-4), Shanghai Shi Feng bio tech ltd；
Poly-l-lysine (MW3-7 ten thousand, A0057-250mg), Shanghai Shi Feng bio tech ltd.
The preparation of cells frozen storing liquid：
Using 100mLRPMI-1640 nutrient solutions as solvent, using dimethyl sulfoxide (DMSO), people's gamma Globulin, poly-l-lysine as
Solute, solvent and solute are mixed.Specially：First 98mLRPMI-1640 nutrient solutions and 2mL dimethyl sulfoxide (DMSO)s are mixed,
People's gamma Globulin, poly-l-lysine are added, people's gamma Globulin, poly-l-lysine final concentration are respectively 10,25 μ g/
mL。
After preparing, it is placed in 4 DEG C of refrigerators and preserves.
Wherein, poly-l-lysine can play a part of preventing people's gamma Globulin from settling.Inventor is specific real at one
Apply in scheme, be also placed in preserving in 4 DEG C of refrigerators, 30 days using the cells frozen storing liquid without poly-l-lysine as a comparison
The concentration of 3 samples measure wherein gamma Globulin is taken according to high uniformity from liquid level to bottom of bottle afterwards, as a result in intermediate altitude sample
The concentration of gamma Globulin is about 5 times of concentration at liquid level, and the concentration of gamma Globulin is about concentration at liquid level in sample at bottom of bottle
11 times.And sedimentation, concentration of 4 DEG C of standings after 6 months at three height is not present in the cell culture fluid for adding poly-l-lysine
Without significant difference.
Certainly, in order to avoid pollution, appropriate antibiotic, such as 50U/mL gentamicins can also be added in cell culture.
Another preparation contrast frozen stock solution is standby, and the contrast frozen stock solution is compared with above-mentioned cells frozen storing liquid only without how poly- L-
Lysine.
Embodiment 2：DCs Multiplying cultures, freeze, recover, anabiosis rate and immunocompetence detection
First, experiment material and method
1st, DCs and CIK preparation
The culture of mononuclearcell：The healthy volunteer's peripheral blood 20ml of taking heparin anti-freezing, it is close with lymphocyte separation medium
Gradient centrifugation (2400 × g, 20min) is spent, the mononuclearcell of gentle aspiration buffy coat is placed in 15ml centrifuge tubes, used
RPMI-1640 (Gibco) medium centrifugal washs 3 times (2000 × g, 5min).Use serum free medium UltraCULTURETM
(Beijing Hui Te is than science and technology) suspension cell, adjustment cell density is 1 × 105/ ml, add Tissue Culture Flask in, put 37 DEG C, 5%
CO2Cultivated in incubator after 2h, jog cell bottle.
The induction of CIK cell：The cell of suspension in above-mentioned cell bottle is drawn onto in another blake bottle, adjustment density is 1
×106/ ml, CIK cell is induced with the UltraCULTURETM containing IL-21000U/ml, CD3-McAb100ng/ml, per 2-
3d half, which is measured, changes liquid, CIK is collected at the 12nd day standby.
The induction of DCs cells：Attached cell in above-mentioned cell bottle is used and contains 100ng/mlhGM-CSF and 1000U/ml
IL-4 UltraCULTURETM continues to cultivate, and 2-3d half, which is measured, changes liquid, to the DCs cells that induction is collected at the 12nd day.
With trypan blue staining living cell counting number.A part of cell is done into killing activity measure, another part cell cryopreservation.
2nd, DCs freezes and recovered
Frozen stock solution of the present invention that DCs is prepared with embodiment 1 respectively and contrast frozen stock solution (face with now match somebody with somebody) be diluted to 3.5 ×
105/ ml, respectively dispenses out 4 pipes and is first placed in -80 DEG C of low temperature refrigerators and cool 12 hours, then is placed in -196 DEG C of liquid nitrogen and preserves.Respectively at
3rd, 6,9 and December it is each it is each from liquid nitrogen take out 1,40 DEG C of water-bath rewarmings are put into rapidly, after suctioning out cell immediately after cell thawing
Suspension, is washed with RPMI-1640 (Gibco) medium centrifugal, and detects DCs anabiosis rates with trypan blue staining.
Trained after DCs recoveries with the UltraCULTURETM serum-frees containing 100ng/mlhGM-CSF and 1000U/mlIL-4
Support base to continue to cultivate 2d, collect DCs standby.
3rd, mtt assay detects DC-CIK killing activity
Cultivate the 12nd day CIK and recovery after be further cultured for 2d DCs by 6:1 ratio is mixed, adjustment cell number be 1 ×
106/ ml, cultivates 4d with the UltraCULTURETM containing IL-2 (1000U/ml), hGM-CSF (100ng/ml), that is, co-cultures
DC-CIK, adjustment density be 4 × 105/ ml, adds in 96 porocyte culture plates containing K562 cells and (imitates target and compare 20:1),
In this, as experimental group.DC-CIK is individually added into addition and is individually added into K562 cells, every group of 5 multiple holes, as a control group.Put 37
DEG C, 5%CO2Continue to cultivate 48h in incubator, supernatant is abandoned in centrifugation, and 20 μ lMTT (5mg/ml) are added per hole, places 37 DEG C, 5%CO2Incubate
4h is cultivated in case, supernatant is abandoned in centrifugation, the μ l of dimethyl sulfoxide (DMSO) 100, vibration dissolving are added per hole.ELIASA 570nm wavelength is surveyed after 15min
D values.DC-CIK killing rate is calculated according to the following formula：
Killing rate (%)=【1- (the independent DC-CIK group D of experimental group D-)/independent K562 cells D】× 100%.
2nd, experimental result
1st, DCs anabiosis rates
The frozen stock solution of the present invention and contrast frozen stock solution prepared using embodiment 1 freezes 3,6,9 to DCs respectively as freezing protective agent
With anabiosis rate in December as shown in table 1 and Fig. 1.With contrasting frozen stock solution ratio, frozen stock solution anabiosis rate of the present invention is higher, freezes protecting effect
More preferably, this explanation is in the presence of no poly-l-lysine, and the DMSO of 2% concentration is not enough to be prevented effectively from cell cryopreservation damage.
Cryopreservation resuscitation rates (%) of the DCs of table 1 behind 3,6,9 and December
| March | June | September | December | |
| Frozen stock solution of the present invention | 97.6±2.3 | 96.8±2.5 | 96.1±2.9 | 95.2±2.6 |
| Contrast frozen stock solution | 91.5±3.7 | 85.7±4.3 | 79.4±3.8 | 70.9±4.1 |
2nd, killing activities of the DC-CIK to K562
Cultivate the CIK of the 12nd day and freeze 3,6,9 and December recovery after be further cultured for 2d DCs co-culture DC-CIK pairs
The killing rate of K562 cells is as shown in table 2 and Fig. 2.With contrast frozen stock solution ratio, killing rate of the frozen stock solution of the present invention to K562 cells
Higher, DCs immunocompetences remain more excellent, and this explanation poly-l-lysine additionally assists in maintaining the immunocompetence for freezing DCs.
Killing rates (%) of the DC-CIK of table 2 to K562
| March | June | September | December | |
| Frozen stock solution of the present invention | 30.9±3.5 | 29.3±3.9 | 29.5±3.6 | 28.9±3.8 |
| Contrast frozen stock solution | 31.7±3.9 | 28.8±4.1 | 23.2±3.6 | 18.5±4.2 |
Above-described embodiment can prove that the cells frozen storing liquid that the present invention is provided is used to that when freezing DCs DCs jelly can be ensured
Anabiosis rate and immunocompetence are deposited, freeze cryopreservation resuscitation rate and immunocompetence in 1 year reduces without obvious；Cells frozen storing liquid of the present invention
In, dimethyl sulfoxide (DMSO) is freezing protective agent, reduces the damage that low temperature is caused to DCs；People's gamma Globulin is cytotostatic agent, can be with
Maintain DCs immunocompetence；Poly-l-lysine is anti-settling agent, it is to avoid storage process people gamma Globulin is heavy to frozen stock solution bottom
Drop, meanwhile, poly-l-lysine has the effect of part freezing protective agent, and the presence of poly-l-lysine causes DMSO body
Product concentration can be reduced to the 1-3% of the present invention from the 10-20% of prior art and still be avoided that the damage that low temperature is caused to DCs
Wound；In addition, poly-l-lysine additionally assists in maintaining DCs immunocompetence.
Above-described embodiment is only used for that technical scheme is explained further, it will be appreciated by those skilled in the art that, appoint
What simple replacement is changed all without departing from the present invention, and protection scope of the present invention is not limited to above-mentioned specific embodiment.
Claims (6)
1. a kind of cells frozen storing liquid based on RPMI-1640 nutrient solutions, adds freezing for effective dose in RPMI-1640 nutrient solutions
Protective agent, cytotostatic agent and anti-settling agent are uniformly mixed, and freezing protective agent is dimethyl sulfoxide (DMSO), it is characterised in that：It is described
Cytotostatic agent is people's gamma Globulin；The anti-settling agent is poly-l-lysine, and the molecular weight ranges of poly-l-lysine are
3-7 ten thousand；The concentration expressed in percentage by volume of the dimethyl sulfoxide (DMSO) is 1-3%.
2. cells frozen storing liquid according to claim 1, it is characterised in that：The mass concentration of people's gamma Globulin is 8-12 μ g/
mL。
3. cells frozen storing liquid according to claim 1, it is characterised in that：The poly-l-lysine is in cells frozen storing liquid
In mass concentration be 20-30 μ g/mL.
4. cells frozen storing liquid according to claim 1, it is characterised in that：Also contain antibiotic in the cells frozen storing liquid.
5. cells frozen storing liquid according to claim 4, it is characterised in that：Antibiotic is gentamicin, and concentration is 50U/mL.
6. any cells frozen storing liquids of claim 1-5 freeze the application of aspect in BMDC.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710635539.8A CN107306939B (en) | 2017-07-31 | 2017-07-31 | Cell cryopreservation liquid for dendritic cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710635539.8A CN107306939B (en) | 2017-07-31 | 2017-07-31 | Cell cryopreservation liquid for dendritic cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107306939A true CN107306939A (en) | 2017-11-03 |
| CN107306939B CN107306939B (en) | 2020-09-15 |
Family
ID=60169931
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710635539.8A Active CN107306939B (en) | 2017-07-31 | 2017-07-31 | Cell cryopreservation liquid for dendritic cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107306939B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110637810A (en) * | 2019-10-18 | 2020-01-03 | 中生康元生物科技(北京)有限公司 | Clinical dendritic cell cryopreservation liquid |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110172315A1 (en) * | 2008-06-27 | 2011-07-14 | Bio Verde. Inc. | Composition for cryopreservation of cells and tissues |
| US20110250632A1 (en) * | 2008-12-18 | 2011-10-13 | Ge Healthcare Uk Limited | Methods for conducting cellular assays |
| US20120258082A1 (en) * | 2009-12-18 | 2012-10-11 | Bavarian Nordic A/S | Production of ifn-lambda by conventional dendritic cells and uses thereof |
| CN104026118A (en) * | 2013-11-13 | 2014-09-10 | 杭州易文赛生物技术有限公司 | Immunization cell frozen stock solution, preparation method and application |
| CN104531617A (en) * | 2015-01-13 | 2015-04-22 | 北京益诺勤生物技术有限公司 | Method for preparing dendritic cells and obtained dendritic cells |
| CN104719282A (en) * | 2015-02-13 | 2015-06-24 | 广州赛莱拉干细胞科技股份有限公司 | Peripheral blood mononuclear cell serum-free freezing medium and freezing method |
| JP2015149905A (en) * | 2014-02-10 | 2015-08-24 | 国立大学法人北陸先端科学技術大学院大学 | Slow vitrification method for cryopreservation of cell sheet and three-dimensional cell culture |
| WO2016063208A1 (en) * | 2014-10-20 | 2016-04-28 | Stempeutics Research Pvt. Ltd. | A cryopreservation composition and methods thereof |
| CN105707058A (en) * | 2016-04-25 | 2016-06-29 | 南京三生生物技术有限公司 | Formula and preparation method of dimethyl-sulfoxide-free protein-free -80 DEG C cell freezing medium |
| CN105994254A (en) * | 2016-07-28 | 2016-10-12 | 广州赛莱拉干细胞科技股份有限公司 | Cryopreservation solution and cryopreservation method of DC cell |
-
2017
- 2017-07-31 CN CN201710635539.8A patent/CN107306939B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110172315A1 (en) * | 2008-06-27 | 2011-07-14 | Bio Verde. Inc. | Composition for cryopreservation of cells and tissues |
| US20110250632A1 (en) * | 2008-12-18 | 2011-10-13 | Ge Healthcare Uk Limited | Methods for conducting cellular assays |
| US20120258082A1 (en) * | 2009-12-18 | 2012-10-11 | Bavarian Nordic A/S | Production of ifn-lambda by conventional dendritic cells and uses thereof |
| CN104026118A (en) * | 2013-11-13 | 2014-09-10 | 杭州易文赛生物技术有限公司 | Immunization cell frozen stock solution, preparation method and application |
| JP2015149905A (en) * | 2014-02-10 | 2015-08-24 | 国立大学法人北陸先端科学技術大学院大学 | Slow vitrification method for cryopreservation of cell sheet and three-dimensional cell culture |
| WO2016063208A1 (en) * | 2014-10-20 | 2016-04-28 | Stempeutics Research Pvt. Ltd. | A cryopreservation composition and methods thereof |
| CN104531617A (en) * | 2015-01-13 | 2015-04-22 | 北京益诺勤生物技术有限公司 | Method for preparing dendritic cells and obtained dendritic cells |
| CN104719282A (en) * | 2015-02-13 | 2015-06-24 | 广州赛莱拉干细胞科技股份有限公司 | Peripheral blood mononuclear cell serum-free freezing medium and freezing method |
| CN105707058A (en) * | 2016-04-25 | 2016-06-29 | 南京三生生物技术有限公司 | Formula and preparation method of dimethyl-sulfoxide-free protein-free -80 DEG C cell freezing medium |
| CN105994254A (en) * | 2016-07-28 | 2016-10-12 | 广州赛莱拉干细胞科技股份有限公司 | Cryopreservation solution and cryopreservation method of DC cell |
Non-Patent Citations (4)
| Title |
|---|
| KAZUAKI MATSUMURA等: "Long-term cryopreservation of human mesenchymal stem cells using carboxylated poly-l-lysine without the addition of proteins or dimethyl sulfoxide", 《JOURNAL OF BIOMATERIALS SCIENCE，POLYMER EDITION》 * |
| 杜国丰: "ε-聚赖氨酸的应用研究进展", 《轻工科技》 * |
| 钟原等: "ε－聚赖氨酸在牛胚胎玻璃化冷冻中的保护作用", 《中国兽医学报》 * |
| 陆士新主编: "《干细胞与肿瘤》", 30 April 2009, 中国协和医科大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110637810A (en) * | 2019-10-18 | 2020-01-03 | 中生康元生物科技(北京)有限公司 | Clinical dendritic cell cryopreservation liquid |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107306939B (en) | 2020-09-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2018023829A1 (en) | Umbilical cord, umbilical cord blood and placenta collection kit | |
| JP2008531051A (en) | Method for providing readily available peripheral blood-derived cellular material and composition thereof | |
| KR20070103431A (en) | Method of providing readily available cellular material derived from cord blood, and a composition thereof | |
| CN108040460A (en) | The method of freezen protective tumor infiltrating lymphocyte | |
| CN109315386A (en) | A kind of frozen stock solution and cryopreservation methods can be used for candidate stem cell or lymphocyte | |
| CN105994254A (en) | Cryopreservation solution and cryopreservation method of DC cell | |
| CN105582004A (en) | Application of chlorogenic acid and/or chlorogenic acid derivative in tumor stem cell treatment drug | |
| CN107581186A (en) | Cells frozen storing liquid and preparation method and application | |
| CN104694473A (en) | Method of automatically extracting immune cells in APB (Adult Peripheral Blood) | |
| CN108029679B (en) | Freezing medium for freezing mononuclear cells | |
| CN104797134B (en) | cell preparation method | |
| CN105754939A (en) | Method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood | |
| CN107306939A (en) | A kind of cells frozen storing liquid for BMDC | |
| CN104789530A (en) | Method for increasing directional differentiation quantity of cord blood megakaryoblasts | |
| CN107711823B (en) | Cell cryopreservation liquid stored at normal temperature and application thereof | |
| CN104472474A (en) | Human adipose tissue-derived stromal cell frozen stock solution | |
| CN106520681A (en) | In vitro culture solution and cryopreservation solution of Ietalurus Punetaus spermatogonial stem cells, and in vitro culture and cryopreservation method | |
| Schrcnemakers et al. | A two-step or equilibrium freezing procedure for the cryopreservation of plant cell suspensions | |
| CN103004751A (en) | Hematopoietic stem cell cryopreserving method and protective agent | |
| CN101188941A (en) | Method and composition for treating diabetes | |
| Zeng et al. | Maintaining viability and characteristics of cholangiocarcinoma tissue by vitrification-based cryopreservation | |
| Stroncek et al. | Stability of cryopreserved white blood cells (WBCs) prepared for donor WBC infusions | |
| CN114190368B (en) | Serum-free immune cell cryopreservation liquid, preparation method and immune cell cryopreservation method | |
| CN114946836A (en) | Micro-tissue serum-free freezing medium and application thereof | |
| Fu et al. | Vitrification of rhesus macaque mesenchymal stem cells and the effects on global gene expression |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Rong Daqian Inventor after: Lei Yunting Inventor after: Hu Panyong Inventor after: Zhang Zhongxiang Inventor after: Hu Wanfu Inventor before: Hu Panyong Inventor before: Zhang Zhongxiang Inventor before: Hu Wanfu |
|
| CB03 | Change of inventor or designer information | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20200825 Address after: Floor 1, building 13, Life Science City, 1777 Dazheng Road, high tech Zone, Jinan City, Shandong Province Applicant after: Shandong Shuifa Life Science Research Co., Ltd Address before: No. 9 Zhongshan Science Park by the road in Liuhe District of Nanjing City, Jiangsu province 211505 Applicant before: NANJING BIOTEKE BIOTECHNOLOGY Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |