CN103070161B - Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) - Google Patents
Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) Download PDFInfo
- Publication number
- CN103070161B CN103070161B CN201310008406.XA CN201310008406A CN103070161B CN 103070161 B CN103070161 B CN 103070161B CN 201310008406 A CN201310008406 A CN 201310008406A CN 103070161 B CN103070161 B CN 103070161B
- Authority
- CN
- China
- Prior art keywords
- cell
- mesenchymal stem
- liquid
- stem cell
- cryopreservation
- 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.)
- Active
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a cryopreservation liquid and a cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC). The cryopreservation liquid is composed of the following materials: 3 to 10 mg/ml of vitamine C, 2 to 8 mg/ml of vitamin E, 2 to 8 mg/ml of lipoic acid, 5 to 20 volume percent of human autoserum, and DMEM-LG culture medium in balancing amount. The cryopreservation liquid for ADSC does not contain animal serum and DMSO, and is composed of human autoserum, lipoic acid, vitamin E, vitamin C and other main materials. A cell cryopreserved with the cryopreservation liquid has the advantages of high palinesthesia rate, no impact on cell growth characteristic, and maintanience of stem cell pluripotency after palinesthesia.
Description
Technical field
The present invention relates to a kind of stem cell cryopreserving liquid, especially a kind of fat mesenchymal stem cell cryopreserving liquid and preparation method thereof.
Background technology
Mescenchymal stem cell (mesenchymal stem cells, MSCs) is the important member of stem cell family, derives from and grows early stage mesoderm and ectoderm.Mescenchymal stem cell finds at first in marrow, has the features such as multi-lineage potential, hematopoiesis support and the implantation of promotion stem cell, immunoregulation and self-replacation and be day by day subject to people's concern because of it.Mescenchymal stem cell is in vivo or under external specific inductive condition, can be divided into the Various Tissues cells such as fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelium, after continuous passage cultivation and freezing preservation, still there is multi-lineage potential, can be used as the injuries of tissues and organs reparation that desirable seed cell causes for old and feeble and pathology.
The mesenchymal stem cell cryopreserving liquid using in prior art contains hyclone and dimethyl sulfoxide (DMSO) (DMSO) more.Hyclone contains multiple growth factor, is usually used in cell in vitro and cultivates, for the adherent of cell or spread over required growth factor is provided on culture matrix; Also can be used for stem cell cryopreserving, the problem that the cytoactive that can slow down declines, can maintain Cell viability for a long time.DMSO, because it has freeze proof effect, is usually used in the frozen of cell.
The hyclone that in prior art, mesenchymal stem cell cryopreserving liquid uses comes from import mostly, and import serum is expensive, and European multinational generation rabid ox disease, and the application of import hyclone has also been subject to certain restriction.There are some researches show that the mescenchymal stem cell contacting with hyclone for a long time can be to the hyclone generation endocytosis in solution medium, likely there is the variation of some protein expression in the mescenchymal stem cell after endocytosis hyclone, after being applied to human body, likely there is the immune response that heterogenous animal albumen causes, thereby cause success rate after mesenchymal stem cell transplantation to reduce and the generation of bad reaction.DMSO has toxicity, can cause damage to freeze-stored cell, and external source animal blood serum has the possibility of pollution to cell, and long-term use affects cytoactive, and follow-up use affects cell clinical treatment and promotes.
Therefore, also a lot of about the research of stem cell cryopreserving liquid.As the publication number Chinese invention patent that is CN101919379, a kind of cryopreserving liquid for freezing and storing umbilical mesenchymal stem cells for long time is disclosed, made by following steps: 1) Cord blood is placed in to aseptic centrifuge tube, centrifugal 15-18min under 1000rpm-1200rpm condition; 2) draw supernatant, be transferred in new centrifuge tube, centrifugal 18-20min under 4000g-4200g condition, supernatant is frozen with umbilical cord blood plasma; 3) get dimethyl sulfoxide (DMSO) and frozenly mix by 1: 9 by volume ratio of umbilical cord blood plasma with above-mentioned, obtain mesenchymal stem cell cryopreserving liquid.In this invention cryopreserving liquid, use the blood plasma of Cord Blood-Derived to substitute hyclone, avoided cell in frozen process, to contact heterogenous animal serum, and reduced cell cryopreservation cost.
Publication number is the Chinese invention patent application of CN102511471, discloses a kind of mesenchymal stem cell cryopreserving liquid that does not contain animal sources albumen and DMSO, is made up of trehalose, carbohydrate-electrolyte solution, ATP and human serum albumins.This cryopreserving liquid has high anabiosis rate (>90%), does not affect the characteristic of stem cell, and cell proliferation does not change, Cell Differentiation does not change; The advantages such as easily use, good stability.
Existing mesenchymal stem cell cryopreserving liquid contains hyclone or DMSO more, even if do not contain hyclone or DMSO, frozen effect is also undesirable; More there is no the cryopreserving liquid about fat mesenchymal stem cell.
Summary of the invention
In view of this, be necessary for the problems referred to above, a kind of fat mesenchymal stem cell cryopreserving liquid and cryopreservation methods that does not contain animal blood serum and DMSO is provided.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
A kind of fat mesenchymal stem cell cryopreserving liquid, is made up of the following component of following content:
As a kind of preferred embodiment of fat mesenchymal stem cell cryopreserving liquid of the present invention, described fat mesenchymal stem cell cryopreserving liquid, is made up of the following component of following content:
A method for frozen fat mesenchymal stem cell, comprises the following steps:
1) aseptic absorption liposuction procedures extraction of lipid liquid, with PBS flushing several, removes medicine used and haemocyte in liposuction procedures; Use type i collagen enzymic digestion, centrifugal, draw upper strata fat and filter.By centrifugal obtained cell suspension, cell precipitation, with after PBS washing, with suspending containing the autoserous DMEM-LG culture fluid of volume fraction 10%, is inoculated in culture dish, is placed in incubator and cultivates after 48 hours, and sucking-off suspension cell liquid, more renews medium; Within 3 days, change liquid 1 time, cultivate when within 10 days, left and right cell reaches more than 80% degree of converging the cultivation of going down to posterity;
2) with pancreatin, cell is digested, discard medium, PBS rinses twice, with trypsin digestion cell, and centrifugal collecting cell;
3) with the cell of the frozen acquisition of fat mesenchymal stem cell cryopreserving liquid of the present invention, be placed on-80 DEG C of refrigerator overnight, within the 2nd day, put into liquid nitrogen container and preserve for a long time.
As a kind of preferred embodiment of the method for the frozen fat mescenchymal stem cell of the present invention fat, described is to digest 1 hour with 37 DEG C of 0.1%I Collagenase Types with type i collagen enzymic digestion.
Compared with prior art, the present invention is a kind of fat mesenchymal stem cell cryopreserving liquid that does not contain animal blood serum and DMSO, by Human autologous serum, lipoic acid, vitamin E, the Main Ingredients and Appearance compositions such as vitamin C, utilize the frozen cell of this cryopreserving liquid to have high anabiosis rate, do not affect cell growth characteristics, the rear cell of recovery keeps the features such as the versatility of stem cell.Without external source animal blood serum, the Human autologous serum of interpolation, lipoic acid, vitamin E and vitamin C do not affect the application of follow-up clinical cell therapy.Human autologous serum, from stem cell cryopreserving person, without foreign protein, and has multiple growth factor, and the ratio of each growth factor conforms to normal ratio in body, after use, cell recovery is had everything to gain and nothing to lose.
Brief description of the drawings
Fig. 1 uses the Growth of Cells form after the frozen cell recovery of different mesenchymal stem cell cryopreserving liquids.Figure 1A is the frozen cell of fat mesenchymal stem cell cryopreserving liquid that uses the embodiment of the present invention 1, and Figure 1B uses the frozen cell of common stem cell cryopreserving liquid.
Fig. 2 uses many differentiation potentials of cell the result after the frozen cell recovery of different mesenchymal stem cell cryopreserving liquids; Fig. 2 A and Fig. 2 C become fat and Osteoblast Differentiation the result after using the frozen cell recovery of the fat mesenchymal stem cell cryopreserving liquid of the embodiment of the present invention 1; Fig. 2 B becomes fat and Osteoblast Differentiation the result after being the frozen cell recovery of the common stem cell cryopreserving liquid of use with Fig. 2 D.
Embodiment
For good explanation the present invention, be described further below in conjunction with specific embodiments and the drawings.
A kind of fat mesenchymal stem cell cryopreserving liquid, is made up of the following component of following content:
Lipoic acid not only tool is water-soluble but also tool is fat-soluble, so it can be deep into each position in cell and play antioxidation, lipoic acid, as a kind of coenzyme, is mutually coordinated sugar, fat, metabolism of amino acid by participating in oxidative deamination reaction, and can be separated removing heavy metals to the murder by poisoning containing sulfydryl enzyme.Also has clinically the effect of anti-fatty liver and reduction cholesterolemia.
Vitamin E is lipovitamin; not only biomembranous 26S Proteasome Structure and Function is had to good protective effect; the vitamin e free radical that Mulberry Extract generates rapidly can further generate non-free radical product with another molecule radical reaction again: fertility quinone, therefore its oxidation resistance is stronger.
Vitamin C, not only as antioxidant protection cell, also participates in hydroxylation reaction, thus with body in collagen generate, proteometabolism, anti-inflammatory, anti-infective, removing toxic substances wait effect directly relevant.
Lipoic acid, vitamin E and vitamin C all have very strong antioxidation activity, can effectively remove the toxic action of interior free yl to cell, thereby play the effect of Cell protection, are applied in cell cryopreservation effectively Cell protection activity.
A method for frozen fat mesenchymal stem cell, comprises the following steps:
1) aseptic absorption liposuction procedures extraction of lipid liquid, with PBS flushing several, removes medicine used and haemocyte in liposuction procedures; Use type i collagen enzymic digestion, centrifugal, draw upper strata fat and filter.By centrifugal obtained cell suspension, cell precipitation, with after PBS washing, with suspending containing the autoserous DMEM-LG culture fluid of volume fraction 10%, is inoculated in culture dish, is placed in 37 DEG C, volume fraction 5%CO
2in incubator, cultivate; After 48 hours, sucking-off suspension cell liquid, more renews medium; Within 3 days, change liquid 1 time, cultivate when within 10 days, left and right cell reaches more than 80% degree of converging the cultivation of going down to posterity;
2) with pancreatin, cell is digested, discard medium, PBS rinses twice, with trypsin digestion cell, and centrifugal collecting cell;
3) with the cell of the frozen acquisition of fat mesenchymal stem cell cryopreserving liquid of the present invention, be placed on-80 DEG C of refrigerator overnight, within the 2nd day, put into liquid nitrogen container and preserve for a long time.
Embodiment 1
A kind of fat mesenchymal stem cell cryopreserving liquid, is made up of the following component of following content:
Be mixed with fat mesenchymal stem cell cryopreserving liquid according to aforementioned proportion and component.Described fat mesenchymal stem cell cryopreserving liquid is by following method freeze-stored cell.
1) the extraction of lipid liquid of aseptic absorption liposuction procedures, with PBS flushing several, removes medicine used and haemocyte in liposuction procedures; With 37 DEG C of digestion of 0.1%I Collagenase Type 1 hour, the centrifugal 5min of 400g/min, drew after the fat of upper strata, mixed with 200 eye mesh screens filtrations; By obtained cell suspension 1200r/min, centrifugal 8min, cell precipitation is with after PBS washing, and adjusting cell concentration is 1 × 10
9individual/L, with suspending containing the autoserous DMEM-LG medium of volume fraction 10%, is inoculated in 10cm culture dish, is placed in 37 DEG C, volume fraction 5%CO
2in incubator, cultivate; After 48 hours, sucking-off suspension cell liquid, more renews medium; Within 3 days, change liquid 1 time, cultivate when within 10 days, left and right cell reaches more than 80% degree of converging the cultivation of going down to posterity;
2) with pancreatin, cell is digested, remove medium in blake bottle or carefully suck the medium in culture plate with rifle, PBS rinses twice, with trypsin digestion cell (gentle as far as possible), centrifugal collecting cell;
3) be mixed with fat mesenchymal stem cell cryopreserving liquid freeze-stored cell, cell density is 1 × 10
6individual/mL, mark freeze-stored cell title after sealing, cryopreserving liquid kind and frozen date; Adopt programmed cooling device to be placed on-80 DEG C of refrigerator overnight, within second day, put into liquid nitrogen container and preserve for a long time.
The present invention is a kind of fat mesenchymal stem cell cryopreserving liquid that does not contain animal blood serum and DMSO, by Human autologous serum, lipoic acid, vitamin E, the Main Ingredients and Appearance compositions such as vitamin C, utilize the frozen cell of this cryopreserving liquid to have high anabiosis rate, do not affect cell growth characteristics, the rear cell of recovery keeps the features such as the versatility of stem cell.Without external source animal blood serum, the Human autologous serum of interpolation, lipoic acid, vitamin E and vitamin C do not affect the application of follow-up clinical cell therapy.Human autologous serum, from stem cell cryopreserving person, without foreign protein, and has multiple growth factor, and the ratio of each growth factor conforms to normal ratio in body, after use, cell recovery is had everything to gain and nothing to lose.
Use common stem cell cryopreserving liquid (surplus is DMEM-LG for hyclone 10%, DMSO10%) in contrast.To the high anabiosis rate of freeze-stored cell, do not affect cell growth characteristics, after recovery, cell keeps the versatility of stem cell to investigate.
Cryopreservation tube is taken out from liquid nitrogen, drop into immediately in 37 DEG C of water-baths, slightly shake.After liquid all melts (general 1-1.5min), take out a small amount of alcohol of spray and be put in superclean bench.Above-mentioned cell suspension is drawn onto in the centrifuge tube of 15mL of dress 10mL medium (with medium, cryopreservation tube is washed one time, the cell being bonded on wall is all washed), 1000 leave heart 5min.Supernatant is outwelled, added 1mL DEME-LG perfect medium, cell is suspended.Be drawn onto shake gently all around in the 10cm culture dish that 10mL medium is housed, the cell in culture dish is uniformly distributed.Mark cell category and date, kinds of culture medium people etc., be put into CO
2in incubator, cultivate, after cell attachment, then trypsinization collecting cell.
Get 50uL cell sap and 50uL platform and expect blue mixing, get a little cell mixture and add haemocytometer, and be beneficial under microscope and calculate cell number.Living cells is not dyeed, and dead cell can be dyed to blueness.Calculate the total cell number of 4 large grid of large nine grids, then remove in 4, then take advantage of in extension rate, then take advantage of in 10
4, be cell number number in every mL.Be every mL cell number=always calculate cell number/4 × extension rate × 10
4.Use after the frozen cell recovery of the fat mesenchymal stem cell cryopreserving liquid of the embodiment of the present invention 1 viable count 2 × 10
6individual/mL, anabiosis rate reaches 90%; Use the frozen cell viable count 1 × 10 of common stem cell cryopreserving liquid
6individual/mL, anabiosis rate is 70%.Use the frozen cell of fat mesenchymal stem cell cryopreserving liquid of the present invention, anabiosis rate is high.
Frozen cell and frozen cell growth characteristics and the versatility of the common stem cell cryopreserving liquid of use of fat mesenchymal stem cell cryopreserving liquid described in observation use embodiment 1 found respectively, the frozen cell of fat mesenchymal stem cell cryopreserving liquid described in use embodiment 1 better maintains the activity of fat stem cell, cell has high anabiosis rate, do not affect cell growth characteristics (seeing Fig. 1), the rear cell of recovery keeps the features such as the versatility of stem cell, and (see Fig. 2, after recovery, cell keeps many differentiation potentials.
The above embodiment has only expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (2)
1. a fat mesenchymal stem cell cryopreserving liquid, is characterized in that, is made up of the following component of following content:
2. fat mesenchymal stem cell cryopreserving liquid according to claim 1, is characterized in that, is made up of the following component of following content:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310008406.XA CN103070161B (en) | 2013-01-09 | 2013-01-09 | Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310008406.XA CN103070161B (en) | 2013-01-09 | 2013-01-09 | Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103070161A CN103070161A (en) | 2013-05-01 |
| CN103070161B true CN103070161B (en) | 2014-08-13 |
Family
ID=48147025
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310008406.XA Active CN103070161B (en) | 2013-01-09 | 2013-01-09 | Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103070161B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2655222C2 (en) * | 2016-05-05 | 2018-05-24 | Константин Валентинович Котенко | Method for adipose tissue preparation for cryopreservation |
Families Citing this family (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105211051B (en) * | 2015-10-29 | 2017-07-07 | 广州赛莱拉干细胞科技股份有限公司 | Cultured NK cell freezing medium and preparation method thereof |
| CN105624114B (en) * | 2016-02-04 | 2017-08-25 | 广东盈生力健康科技有限公司 | A kind of stem cell differential medium and its application |
| CN105532649B (en) * | 2016-02-16 | 2016-09-21 | 河北佑仁生物科技有限公司 | A kind of frozen stock solution preserved for a long time for adult stem cell and preparation method thereof |
| CN106222134A (en) * | 2016-07-29 | 2016-12-14 | 中卫华医（北京）生物科技有限公司 | The cultural method of effective acquisition fat mesenchymal stem cell |
| CN106754688A (en) * | 2016-11-21 | 2017-05-31 | 深圳市衍生生物科技有限公司 | A kind of efficient method for resuscitation for freezing PMNC |
| CN108285888A (en) * | 2017-01-10 | 2018-07-17 | 刘金宏 | The preparation method of fresh mescenchymal stem cell |
| CN106947734A (en) * | 2017-02-28 | 2017-07-14 | 兰赫(上海)生物科技有限公司 | A kind of Endometrial stem cell enrichment procedure and its application |
| CN108849859B (en) * | 2018-07-27 | 2020-12-01 | 武汉大学 | Cryopreservation solution for leukemia cell sample and preparation method thereof |
| CN109090102A (en) * | 2018-09-05 | 2018-12-28 | 成都汇欣生命科技有限公司 | A kind of general serum-free frozen stock solution of mescenchymal stem cell and preparation method thereof |
| CN109566600A (en) * | 2018-12-29 | 2019-04-05 | 中国医学科学院整形外科医院 | It is a kind of to freeze pre-treating method for fat stem cell |
| CN110012897A (en) * | 2019-03-11 | 2019-07-16 | 广州赛莱拉干细胞科技股份有限公司 | Stem cell medicine and its application in the drug of preparation treatment osteoarthritis |
| CN110839614B (en) * | 2019-11-27 | 2022-02-11 | 西南医科大学 | Autologous umbilical cord mesenchymal stem cell cryopreservation liquid and preparation and cryopreservation methods thereof |
| 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 |
| CN112075418B (en) * | 2020-09-29 | 2021-10-22 | 惟力维斯(上海)医学科技有限公司 | Adipose mesenchymal stem cell cryopreservation liquid and adipose mesenchymal stem cell cryopreservation method |
| CN112075417B (en) * | 2020-09-30 | 2021-10-08 | 广东赛尔生物科技有限公司 | Adipose mesenchymal stem cell cryopreservation liquid and cryopreservation method thereof |
| CN112841173A (en) * | 2021-01-25 | 2021-05-28 | 赵丽歌 | Hematopoietic stem cell cryopreservation liquid and cryopreservation method |
| CN112841176A (en) * | 2021-01-28 | 2021-05-28 | 暨南大学 | Long-term cryopreservation medium for adipose-derived stem cells and preparation method and application thereof |
| CN113455498A (en) * | 2021-08-12 | 2021-10-01 | 郑州贝因生物科技有限公司 | Adipose mesenchymal stem cell cryopreservation liquid and cryopreservation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011083472A2 (en) * | 2010-01-07 | 2011-07-14 | Core Dynamics Ltd. | Method for preserving cancellous bone samples and preserved cancellous bone tissue |
| CN102349500A (en) * | 2011-11-10 | 2012-02-15 | 成都清科生物科技有限公司 | Mesenchymal stem cell self-preserving liquid |
-
2013
- 2013-01-09 CN CN201310008406.XA patent/CN103070161B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011083472A2 (en) * | 2010-01-07 | 2011-07-14 | Core Dynamics Ltd. | Method for preserving cancellous bone samples and preserved cancellous bone tissue |
| CN102349500A (en) * | 2011-11-10 | 2012-02-15 | 成都清科生物科技有限公司 | Mesenchymal stem cell self-preserving liquid |
Non-Patent Citations (4)
| Title |
|---|
| 人脂肪间充质干细胞的分离培养与生物学特性;毕薇薇 等;《中国组织工程研究与临床康复》;20111001;第15卷(第40期);参见第7494页实验方法，人脂肪间充质干细胞的分离、培养，脂肪间充质干细胞表面抗原标志的测定，脂肪间充质干细胞的冻存与复苏 * |
| 成人脂肪间充质干细胞冻存前后的生物学特性;樊艳 等;《中国组织工程研究与临床康复》;20080617;第12卷(第25期);参见摘要 * |
| 樊艳 等.成人脂肪间充质干细胞冻存前后的生物学特性.《中国组织工程研究与临床康复》.2008,第12卷(第25期), |
| 毕薇薇 等.人脂肪间充质干细胞的分离培养与生物学特性.《中国组织工程研究与临床康复》.2011,第15卷(第40期), |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2655222C2 (en) * | 2016-05-05 | 2018-05-24 | Константин Валентинович Котенко | Method for adipose tissue preparation for cryopreservation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103070161A (en) | 2013-05-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103070161B (en) | Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) | |
| CN102511471B (en) | Mesenchymal stem cell frozen stock solution and preparation method thereof | |
| JP5341059B2 (en) | Stem cell suspension | |
| CN103583511B (en) | A kind of mesenchymal stem cell cryopreserving liquid and parenteral solution | |
| CN102660497B (en) | Method for freezing and reviving umbilical cord tissues and for separating and increasing stem cells | |
| CN102807966B (en) | Method for freezing and thawing placental whole cells and separating and expanding stem cells | |
| CN106465710B (en) | Adipose tissue cryopreservation liquid and adipose tissue cryopreservation method | |
| CN102660502B (en) | Methods for freezing and thawing whole cell of umbilical cord and separating and augmenting stem cell | |
| US10104881B2 (en) | Composition comprising plant-derived recombinant human serum albumin, lipids, and plant protein hydrolysates as active ingredients for cryopreservation of stem cells or primary cells | |
| JP7401865B2 (en) | Methods for obtaining enriched populations of functional mesenchymal stem cells, cells obtained thereby, and compositions comprising the cells | |
| KR101321144B1 (en) | Vitrification medium for stem cells and vitrification method for stem cells using the same | |
| CN102660501A (en) | Method for separating and amplifying mesenchymal stem cell from fresh tissue of umbilical cord | |
| CN107494517A (en) | Serum-free frozen stock solution and its application in mescenchymal stem cell is frozen | |
| CN105534848A (en) | Cosmetic or pharmaceutical composition and application thereof | |
| CN108056095A (en) | A kind of fat mesenchymal stem cell transport protection liquid and its application | |
| CN101974486A (en) | Method for extracting mesenchymal stem cells from trace human fatty tissues and massively culturing | |
| CN108456657A (en) | Dog umbilical cord mesenchymal stem cells and preparation method thereof and cryopreservation methods | |
| KR20100084620A (en) | Cell composition for tissue regeneration | |
| CN108057014A (en) | A kind of preparation method of the stem cell medicine of beauty and skin care | |
| CN106614524B (en) | Preservation solution and preservation method for mesenchymal stem cells | |
| CN102965337A (en) | Method for separating and extracting human subcutaneous adipose-derived mesenchymal stem cells and special culture medium for extraction | |
| CN106701670A (en) | Methods for enhancing bioactive factor secretion capacity of mesenchymal stem cells and extracting active factors in culture solution | |
| CN110317782A (en) | Improve the method survived after mescenchymal stem cell recovery and frozen stock solution used | |
| CN104771414A (en) | Adipose-derived stem cell preparation and preparation method thereof | |
| CN110317781A (en) | The method of mesenchymal stem cell cryopreserving and recovery |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: FENG WENFENG Free format text: FORMER OWNER: GUANGZHOU ETHICAL BIOTECHNOLOGY CO., LTD. Effective date: 20140428 Owner name: SHE ZHIYONG CHEN JIANXING FANG HAIQING Effective date: 20140428 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| C53 | Correction of patent for invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Feng Wenfeng Inventor after: She Zhiyong Inventor after: Fang Haiqing Inventor after: Chen Jianxing Inventor before: Feng Wenfeng Inventor before: She Zhiyong Inventor before: Fang Haiqing |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 510070 GUANGZHOU, GUANGDONG PROVINCE TO: 510000 GUANGZHOU, GUANGDONG PROVINCE Free format text: CORRECT: INVENTOR; FROM: FENG WENFENG SHE ZHIYONG FANG HAIQING TO: FENG WENFENG SHE ZHIYONG FANG HAIQING CHEN JIANXING |
|
| TA01 | Transfer of patent application right |
Effective date of registration: 20140428 Address after: 510000 Guangdong city of Guangzhou province Haizhuqu District Xiaogang Road No. 74 room 503 Applicant after: Feng Wenfeng Applicant after: She Zhiyong Applicant after: Chen Jianxing Applicant after: Fang Haiqing Address before: Guangzhou City, Guangdong province 510070 martyrs Road No. 76 Kiu building 13 Building G room Applicant before: Guangzhou Ethical Biotech Co., Ltd. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |