CN102428911A - Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block - Google Patents
Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block Download PDFInfo
- Publication number
- CN102428911A CN102428911A CN2011103434259A CN201110343425A CN102428911A CN 102428911 A CN102428911 A CN 102428911A CN 2011103434259 A CN2011103434259 A CN 2011103434259A CN 201110343425 A CN201110343425 A CN 201110343425A CN 102428911 A CN102428911 A CN 102428911A
- Authority
- CN
- China
- Prior art keywords
- protection liquid
- umbilical cord
- cryogenic
- liquid
- 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.)
- Pending
Links
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a cryogenic storing protection liquid for a human umbilical cord Wharton's jelly tissue block. One liter of cryogenic storing protection liquid comprises 1-1.4 mol of permeable cryoprotectant, 0.1-0.5 mol of impermeable cryoprotectant and the balance of base liquid. The cryogenic storing protection liquid provided by the invention contains the permeable cryoprotectant and the impermeable cryoprotectant at the same time, the concentration of the permeable cryoprotectant is low, so that good cryogenic storage effect can be achieved, and the toxicity of cells is also small. Meanwhile, the cryogenic storing protection liquid contains the base liquid, not only can nutrients be provided for the human umbilical cord Wharton's jelly tissue block, but also the toxicity of cryoprotectants on cells in the Wharton's jelly tissue block is lowered. The human umbilical cord Wharton's jelly tissue block cryogenically stored by the cryogenic storing protection liquid can be used for preparing high quality umbilical cord mesenchymal stem cells, few cells are damaged, and the cryogenic storing protection liquid has small residue, small toxic and side effects and no harm to human bodies in clinic use.
Description
Technical field
The present invention relates to the frozen protection liquid of people's umbilical cord China Tong Shi glue piece of tissue.
Background technology
Cell in people's umbilical cord China Tong Shi glue (Wharton's Jelly) has can self and the characteristic of stem cell such as multidirectional differentiation, multiplication capacity be strong, and we are referred to as umbilical cord mesenchymal stem cells (huMSCs).Mescenchymal stem cell is the important member of stem cell family, is a kind of stem cell of reduced immunogenicity, have hematopoiesis support and promote characteristics such as stem cell is implanted, regulation and control immunity, and be to be used for the desirable seed cell that injuries of tissues and organs old and feeble and that pathology causes is repaired.Umbilical cord mesenchymal stem cells is prone to obtain, and has all characteristics of mescenchymal stem cell because of its wide material sources, receives people's attention day by day, and increasing people begins to store umbilical cord mesenchymal stem cells and thinks the possible period of want or need in the future.The prescription of frozen protection liquid and frozen operation scheme directly influence the quality of mescenchymal stem cell in umbilical cord China Tong Shi glue of storage, and be most important to cell transplantation effect in the future.Cell or tissue freezing are preserved several critical links: the selection of cryoprotector and combination, the proportioning of various cryoprotectors, frozen step, frozen temperature and recovery temperature.
At present; The traditional cell freezing method of the many employings of the frozen method of umbilical cord mesenchymal stem cells, cell freezing mainly adopts the frozen protection liquid that contains the permeability cryoprotector, and effect is to reduce the cell freezing point; Reduce the formation of ice crystal, alleviate cells injury in the refrigerating process.Generally; People select the single frozen stem cell of permeability cryoprotector dimethyl sulfoxide (DMSO); Have the residual of frozen protection liquid after the recovery of frozen back; The stem cell of the residual frozen protection liquid dimethyl sulfoxide (DMSO) of clinical use has higher toxic and side effect, is prone to cause bad reactions such as nauseating, vomiting, stomachache.
Summary of the invention
The objective of the invention is to adopt by what basal liquid, permeability cryoprotector, impermeability cryoprotector were formed and directly people's umbilical cord China Tong Shi glue piece of tissue is carried out freezing frozen protection liquid; Frozen effective; Cellular damage is few; Frozen protection liquid is residual low, and toxic and side effect is little, during clinical use human body is not had injury.
In order to achieve the above object, the frozen protection liquid of umbilical cord China's Tong Shi glue piece of tissue of the present invention, the frozen protection liquid of 1L comprises permeability cryoprotector 1-1.4 mol, impermeability cryoprotector 0.1-0.5mol, surplus is a basal liquid, wherein
Said basal liquid comprises culture fluid DMEM/F12, phosphate buffer PBS, physiological saline, contains the culture fluid DMEM/F12 of 10% hyclone, from body bleeding of the umbilicus serum, the STEMPRO of Invitrogen company
MSC SFM culture fluid, the Stem Cell MesenCult of company
The combination of one or more the in-XF Medium culture fluid;
Said permeability cryoprotector comprises one or more the combination in dimethyl sulfoxide (DMSO), ethylene glycol, propane diols, the glycerine;
Said impermeability cryoprotector comprises one or more the combination in sucrose, mannitol, the trehalose.
Compared with prior art; What frozen protection liquid of the present invention was frozen is the umbilical cord China Tong Shi glue piece of tissue that comprises mescenchymal stem cell; Frozen protection formula of liquid is neither the frozen protection liquid that contains single cryoprotector of frozen stem cell; Neither contain the glass frozen preservation protection liquid of high concentration cryoprotector by the cryopreserved tissue piece, but adopt the frozen protection liquid of forming by permeability cryoprotector and impermeability cryoprotector, the basal liquid of low concentration.Adopted the culture fluid DMEM/F12 that comprises culture fluid DMEM/F12, contain 10% hyclone in this frozen protection liquid, from the basal liquid of body bleeding of the umbilicus serum, phosphate buffer PBS etc.; Can guarantee the osmotic equilibrium that cell is inside and outside; Especially having adopted what come from same individuality is that piece of tissue provides nutrient from body bleeding of the umbilicus serum as basal liquid; Be simple and easy to from body bleeding of the umbilicus serum, clinical easy realization do not have the adding of outer derived components; The environment that has guaranteed piece of tissue inner cell in the frozen process to greatest extent is consistent with internal milieu, and is frozen effective; Adopted the permeability cryoprotector that comprises dimethyl sulfoxide (DMSO), ethylene glycol, propane diols, glycerine etc.; Can permeate through cell membranes be penetrated in the cell; Reduce the formation of ice crystal in cell when freezing, and concentration is in all on the lower level, reduced the toxicity of pair cell to greatest extent; Adopted the impermeability cryoprotector that comprises sucrose, mannitol, trehalose etc.; Molecular weight is big; A part seldom just can increase the viscosity of solution greatly, and the inside and outside osmotic pressure of cell membrane makes cell dehydration when alleviating cell cryopreservation; The ice crystal that reduces cell when freezing forms, and to the stem cell nontoxic property in the piece of tissue.
[0007] the present invention adopts the direct frozen protection liquid of people's umbilical cord China Tong Shi glue piece of tissue; Considered to reduce the toxicity of the residual pair cell of frozen protection liquid simultaneously in piece of tissue recovery back; The quality of the cell of turning out behind the assurance piece of tissue cryopreservation resuscitation can be for providing the umbilical cord mesenchymal stem cells of better quality clinically.
Description of drawings
Fig. 1 is for freezing the umbilical cord China Tong Shi glue piece of tissue of preserving 30 days through this frozen protection liquid cooling, the P0 that turns out is for human umbilical cord mesenchymal stem cells growth conditions figure;
Fig. 2 is for freezing the umbilical cord China Tong Shi glue piece of tissue of preserving 30 days through this frozen protection liquid cooling, the P1 that turns out induces evaluation figure for the one-tenth fat of human umbilical cord mesenchymal stem cells;
Fig. 3 is for freezing the umbilical cord China Tong Shi glue piece of tissue of preserving 30 days through this frozen protection liquid cooling, the P1 that turns out is for the osteogenic induction evaluation figure of human umbilical cord mesenchymal stem cells.
Embodiment
To the detailed description of the invention, but protection scope of the present invention is not limited to the scope that embodiment lists through following examples.
Instance one:
Get the phosphate buffer PBS of 400ml; Filter with the 0.22um filter membrane behind the abundant mixing of adding trehalose 0.5mol filters back 4 ℃ of precoolings; Add the ethylene glycol 1mol from body bleeding of the umbilicus serum 100ml, 4 ℃ of precoolings of 4 ℃ of precoolings, the dimethyl sulfoxide (DMSO) 0.4mol of 4 ℃ of precoolings on the ice bath successively, add phosphate buffer PBS to 1L and promptly be the frozen protection liquid of people's umbilical cord China's Tong Shi glue piece of tissue.
Instance two:
Get the culture fluid DMEM/F12 of 400ml; Filter with the 0.22um filter membrane behind the abundant mixing of adding trehalose 0.1mol filters back 4 ℃ of precoolings; Add the dimethyl sulfoxide (DMSO) 0.1mol from body bleeding of the umbilicus serum 200ml, 4 ℃ of precoolings of 4 ℃ of precoolings, the ethylene glycol 1mol of 4 ℃ of precoolings on the ice bath successively, add culture fluid DMEM/F12 to 1L and promptly be the frozen protection liquid of people's umbilical cord China's Tong Shi glue piece of tissue.
Instance three:
Get the phosphate buffer PBS of 500ml; Filter with the 0.22um filter membrane behind the abundant mixing of adding mannitol 0.2mol filters back 4 ℃ of precoolings; The dimethyl sulfoxide (DMSO) 1mol that adds 4 ℃ of precoolings on the ice bath adds phosphate buffer PBS to 1L and promptly is the frozen protection liquid of people's umbilical cord China's Tong Shi glue piece of tissue.
Instance four:
Get the culture fluid DMEM/F12 that contains 10% hyclone of 500ml; Filter with the 0.22um filter membrane behind the abundant mixing of adding sucrose 0.2mol filters back 4 ℃ of precoolings; Add the ethylene glycol 1.1mol of 4 ℃ of precoolings, the propane diols 0.2mol of 4 ℃ of precoolings on the ice bath successively, add the culture fluid DMEM/F12 to 1L that contains 10% hyclone and promptly be the frozen protection liquid of people's umbilical cord China's Tong Shi glue piece of tissue.
Instance five:
Get part culture fluid DMEM/F12 of 400ml; Filter with the 0.22um filter membrane behind the abundant mixing of adding trehalose 0.2mol filters back 4 ℃ of precoolings; Add the dimethyl sulfoxide (DMSO) 1.1mol from body bleeding of the umbilicus serum 100ml, 4 ℃ of precoolings of 4 ℃ of precoolings, the glycerine 0.2mol of 4 ℃ of precoolings on the ice bath successively, add culture fluid DMEM/F12 to 1L and promptly be the frozen protection liquid of people's umbilical cord China's Tong Shi glue piece of tissue.
Instance six:
Get the STEMPRO of the Invitrogen company of 400ml
MSC SFM culture fluid; Filter with the 0.22um filter membrane behind the abundant mixing of adding mannitol 0.3mol filters back 4 ℃ of precoolings; Add the ethylene glycol 0.8mol of the physiological saline 100ml of 4 ℃ of precoolings, 4 ℃ of precoolings, the dimethyl sulfoxide (DMSO) 0.3mol of 4 ℃ of precoolings on the ice bath successively, add the STEMPRO of Invitrogen company
MSC SFM culture fluid to 1L promptly be the frozen protection liquid of people's umbilical cord China Tong Shi glue piece of tissue.
Instance seven:
Get the STEMPRO of Invitrogen company of 200ml
MSC SFM culture fluid adds the 400ml Stem Cell MesenCult of company
-XF Medium culture fluid, the filter with the 0.22um filter membrane behind the abundant mixing of adding trehalose 0.3mol filters back 4 ℃ of precoolings, adds the ethylene glycol 1mol of 4 ℃ of precoolings, the propane diols 0.4mol of 4 ℃ of precoolings on the ice bath successively, adds the STEMPRO of Invitrogen company
MSC SFM culture fluid to 1L promptly be the frozen protection liquid of people's umbilical cord China Tong Shi glue piece of tissue.
After the frozen people's umbilical cord China Tong Shi glue piece of tissue of the frozen protection liquid of such scheme preparation is recovered; Cell growth condition is good; Cell growth curve is no abnormal, becomes the differentiation capability of inducing of fat, skeletonization not lose the cell surface marker indifference that surface markers and fresh piece of tissue are turned out.
Below be that the test of preserving umbilical cord China Tong Shi glue effect is frozen in the frozen protection liquid cooling of preferred example one:
Freeze the umbilical cord China Tong Shi glue piece of tissue of preserving 30 days through this frozen protection liquid cooling, cultivate after 15 days, P0 adds up to 1.74 x10 for human umbilical cord mesenchymal stem cells
5/ gram, the cytomixis in P0 generation is placed under the inverted microscope and takes pictures to 80-90%, and cell shape is short fusiformis, big or small homogeneous, marshalling, as shown in Figure 1.
Freeze the umbilical cord China Tong Shi glue piece of tissue of preserving 30 days through this frozen protection liquid cooling, turn out P1 for human umbilical cord mesenchymal stem cells, with 1x10
4The cell concentration of/ml is inoculated in 24 well culture plates, the 0.5ml/ hole; Cytomixis is to 60-80%, and the one-tenth fat that changes Invitrogen company into is induced perfect medium, the 0.5ml/ hole; Full dose was changed inducing culture liquid in every 3-4 days; After inducing 14 days, take pictures behind the oil red O stain, have fat to drip generation in the visible cell, as shown in Figure 2.
Freeze the umbilical cord China Tong Shi glue piece of tissue of preserving 30 days through this frozen protection liquid cooling, the P1 that cultivation grows is for human umbilical cord mesenchymal stem cells, with 1x10
4The cell concentration of/ml is inoculated in 24 well culture plates, the 0.5ml/ hole; Cytomixis changes the osteogenic induction perfect medium of Invitrogen company into, the 0.5ml/ hole to 60-80%; Full dose was changed inducing culture liquid in every 3-4 days; After inducing 7 days, take pictures after the alizarin red dyeing, visible have a large amount of calcium tubercles to produce, as shown in Figure 3.
Claims (1)
1. the frozen protection liquid of umbilical cord China's Tong Shi glue piece of tissue, the frozen protection liquid of 1L comprises permeability cryoprotector 1-1.4 mol, impermeability cryoprotector 0.1-0.5mol, surplus is a basal liquid;
Said basal liquid comprises culture fluid DMEM/F12, phosphate buffer PBS, physiological saline, the culture fluid DMEM/F12 that contains 10% hyclone, bleeding of the umbilicus serum, the STEMPRO of Invitrogen company
MSC SFM culture fluid, the Stem Cell MesenCult of company
The combination of one or more the in-XF Medium culture fluid;
Said permeability cryoprotector comprises one or more the combination in dimethyl sulfoxide (DMSO), ethylene glycol, propane diols, the glycerine;
Said impermeability cryoprotector comprises one or more the combination in sucrose, mannitol, the trehalose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103434259A CN102428911A (en) | 2010-11-03 | 2011-11-03 | Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105287211A CN101971798A (en) | 2010-11-03 | 2010-11-03 | Human umbilical cord Wharton jelly tissue block freezing protection liquid |
| CN201010528721.1 | 2010-11-03 | ||
| CN2011103434259A CN102428911A (en) | 2010-11-03 | 2011-11-03 | Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102428911A true CN102428911A (en) | 2012-05-02 |
Family
ID=43571734
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010105287211A Pending CN101971798A (en) | 2010-11-03 | 2010-11-03 | Human umbilical cord Wharton jelly tissue block freezing protection liquid |
| CN2011103434259A Pending CN102428911A (en) | 2010-11-03 | 2011-11-03 | Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010105287211A Pending CN101971798A (en) | 2010-11-03 | 2010-11-03 | Human umbilical cord Wharton jelly tissue block freezing protection liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN101971798A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105961374A (en) * | 2016-07-04 | 2016-09-28 | 深圳市合康生物科技股份有限公司 | Cell cryopreservation fluid |
| CN106857498A (en) * | 2016-12-27 | 2017-06-20 | 科兴（大连）疫苗技术有限公司 | A kind of cell cryopreservation protection liquid and its application without dimethyl sulfoxide (DMSO) |
| CN114982744A (en) * | 2022-07-07 | 2022-09-02 | 广州沙艾生物科技有限公司 | Umbilical cord stem cell storage solution and preparation method thereof |
| WO2023164799A1 (en) * | 2022-03-01 | 2023-09-07 | 傅毓秀 | Wharton's jelly product capable of promoting osteogenesis |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102511471B (en) * | 2011-12-15 | 2013-08-28 | 成都清科生物科技有限公司 | Mesenchymal stem cell frozen stock solution and preparation method thereof |
| CN102763639B (en) * | 2012-07-19 | 2014-01-22 | 童亚林 | Composite anti-freezing liquid and application thereof, and method for preserving human amnion by using same |
| CN104145943B (en) * | 2014-07-21 | 2016-02-17 | 重庆市红汇脐血干细胞中心有限公司 | A kind of frozen protection liquid of people's umbilical cord China Tong Shi glue tissue and preparation and application thereof |
| CN104396940B (en) * | 2014-10-11 | 2016-07-13 | 张炳强 | A kind of tissue samples preserves liquid and preparation method thereof |
| CN104472473B (en) * | 2014-11-20 | 2016-03-16 | 黑龙江天晴干细胞股份有限公司 | Umbilical cord tissue block cryopreservation methods |
| CN106508892A (en) * | 2016-11-30 | 2017-03-22 | 广州赛莱拉干细胞科技股份有限公司 | Umbilical cord tissue cryopreservation solution and cryopreservation method |
| CN106417260A (en) * | 2016-11-30 | 2017-02-22 | 广州赛莱拉干细胞科技股份有限公司 | Umbilical cord tissue cryoprotectant |
| CN107996558A (en) * | 2017-12-21 | 2018-05-08 | 湖南丰晖生物科技有限公司 | Cells frozen storing liquid and its application |
| CN108782544B (en) * | 2018-06-28 | 2021-01-01 | 吉林省拓华生物科技有限公司 | Cryopreservation solution and method for cryopreservation of large amount of mesenchymal stem cells |
| US20210315199A1 (en) * | 2018-08-10 | 2021-10-14 | Predictive Technology Group, Inc. | Cryopreservation medium for umbilical cord mscs derived from wharton's jelly |
| CN109122665A (en) * | 2018-09-12 | 2019-01-04 | 银丰生物工程集团有限公司 | With the method for a variety of stem cells of freezing and storing umbilical tissue preserration, method for resuscitation and application |
| CN111011363B (en) * | 2019-12-16 | 2021-12-17 | 广东唯泰生物科技有限公司 | Mesenchymal stem cell cryopreservation liquid, cryopreservation method, preservation kit and recovery method |
| CN112438252B (en) * | 2020-11-20 | 2021-06-08 | 上海欧易生物医学科技有限公司 | Tissue freezing and thawing method suitable for mouse heart tissue single cell sequencing |
| CN114467913B (en) * | 2021-11-22 | 2022-11-11 | 武汉赛尔朗灵科技有限公司 | Long-term cryopreservation and recovery method for animal tumor tissue, clinical tumor tissue and biopsy tumor tissue sample |
| CN116138247A (en) * | 2023-01-31 | 2023-05-23 | 清华大学 | Combined reagent for animal tissue cryoprotection and application and method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101210232A (en) * | 2006-12-28 | 2008-07-02 | 天津昂赛细胞基因工程有限公司 | Mesenchyme stem cell preserving fluid and use thereof |
-
2010
- 2010-11-03 CN CN2010105287211A patent/CN101971798A/en active Pending
-
2011
- 2011-11-03 CN CN2011103434259A patent/CN102428911A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101210232A (en) * | 2006-12-28 | 2008-07-02 | 天津昂赛细胞基因工程有限公司 | Mesenchyme stem cell preserving fluid and use thereof |
Non-Patent Citations (3)
| Title |
|---|
| JACOB HANNA 等: "Preservation of stem cells", 《ORGANOGENESIS》 * |
| MOON 等: "Successful vitrification of human amnion-derived mesenchymal stem cells", 《HUMAN REPRODUCTION》 * |
| 王有为 等: "人脐带间充质干细胞冻存复苏后的生物学特征", 《中国组织工程研究与临床康复》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105961374A (en) * | 2016-07-04 | 2016-09-28 | 深圳市合康生物科技股份有限公司 | Cell cryopreservation fluid |
| CN106857498A (en) * | 2016-12-27 | 2017-06-20 | 科兴（大连）疫苗技术有限公司 | A kind of cell cryopreservation protection liquid and its application without dimethyl sulfoxide (DMSO) |
| CN106857498B (en) * | 2016-12-27 | 2021-05-14 | 科兴（大连）疫苗技术有限公司 | Cell cryopreservation protective solution without dimethyl sulfoxide and application thereof |
| WO2023164799A1 (en) * | 2022-03-01 | 2023-09-07 | 傅毓秀 | Wharton's jelly product capable of promoting osteogenesis |
| CN114982744A (en) * | 2022-07-07 | 2022-09-02 | 广州沙艾生物科技有限公司 | Umbilical cord stem cell storage solution and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101971798A (en) | 2011-02-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102428911A (en) | Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block | |
| CN102428910A (en) | Protecting liquid for freezing and storing human umbilical cord Wharton jelly tissue block | |
| CN107027743B (en) | Cell cryopreservation solution and cell cryopreservation method | |
| Weng et al. | Dimethyl sulfoxide-free cryopreservation for cell therapy: A review | |
| CN101720753B (en) | Cryopreservation solution of tissue engineering products and application method thereof | |
| CN108207930B (en) | Cocktail type cryoprotectant and application thereof | |
| CN104145943B (en) | A kind of frozen protection liquid of people's umbilical cord China Tong Shi glue tissue and preparation and application thereof | |
| CN101974484B (en) | Method for preparing human umbilical cord mesenchymal stem cells | |
| CN109329271A (en) | A kind of clinical mescenchymal stem cell freezen protective liquid and preparation method thereof | |
| CN105961374A (en) | Cell cryopreservation fluid | |
| CN102640745B (en) | Ultralow-temperature cryopreservation and recovery method for embryonic materials | |
| CN107156108A (en) | A kind of peripheral hematopoietic stem cells preserve liquid and preparation method | |
| CN104396940A (en) | Tissue sample preservative solution and preparation method thereof | |
| CN106465710A (en) | Fatty tissue frozen stock solution and fatty tissue cryopreservation methods | |
| CN106332869A (en) | Adipose mesenchymal stem cell cryoprotectant and method for cryopreservation of adipose mesenchymal stem cells | |
| CN110432259A (en) | A kind of frozen solution and the cells frozen storing liquid containing it and its application in cell cryopreservation | |
| CN105211052B (en) | Frozen stock solution of cultured NKT cells and preparation method thereof | |
| CN106942200A (en) | One kind freezes protection liquid and its application | |
| CN102763642A (en) | Cryoprotectant and method for cryopreserving placenta amnion and chorion | |
| CN113925049B (en) | Cell preservation solution for maintaining cell activity and preparation method and application thereof | |
| CN101070534A (en) | Method for freezing preservation nervous stem cells | |
| CN106614526A (en) | Vitrified cryopreservation method for cartilage | |
| CN102144629A (en) | Organ cryopreservation fluid | |
| CN109221089A (en) | A kind of method that the frozen stock solution and adipose tissue of adipose tissue freeze | |
| CN104450521A (en) | Method for preserving cordyceps militaris strain for long time |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120502 |