CN112471138B - Universal preservation solution for human umbilical cord, amnion and placenta samples and preparation method thereof - Google Patents
Universal preservation solution for human umbilical cord, amnion and placenta samples and preparation method thereof Download PDFInfo
- Publication number
- CN112471138B CN112471138B CN202011438054.8A CN202011438054A CN112471138B CN 112471138 B CN112471138 B CN 112471138B CN 202011438054 A CN202011438054 A CN 202011438054A CN 112471138 B CN112471138 B CN 112471138B
- Authority
- CN
- China
- Prior art keywords
- preservation solution
- universal
- stem cells
- mesenchymal stem
- concentration
- 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
Images
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
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- 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
- A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts
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)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of stem cells, and particularly relates to a universal preservation solution for human umbilical cord, amnion and placenta samples and a preparation method thereof. In the universal preservation solution, a DMEM basic culture medium provides the most basic nutrition for mesenchymal stem cells of a sample; the insulin can promote the mesenchymal stem cells of the sample to absorb and utilize sugar in the universal preservation solution, and maintain the activity of the cells; the human serum albumin provides dynamic balance for the mesenchymal stem cells and maintains the osmotic pressure of the cells; the vitamin C can prevent the damage of free radicals generated by long-time oxidation to the mesenchymal stem cells; EGF can promote the proliferation of mesenchymal stem cells and maintain the original form of the cells. Experimental results show that the preservation solution can be used for preserving umbilical cords, amnion and placenta for a long time, the obtained mesenchymal stem cells are large in quantity and high in survival rate, and human serum albumin, insulin, vitamin C and EGF have a synergistic effect in improving the number of the mesenchymal stem cells in the preserved umbilical cords and the survival rate of the mesenchymal stem cells.
Description
Technical Field
The invention belongs to the technical field of stem cells, and particularly relates to a universal preservation solution for human umbilical cord, amnion and placenta samples and a preparation method thereof.
Background
Mesenchymal stem cells are cells with self-renewal and multipotential differentiation potential, also called multipotential stromal cells, MSCs for short, belonging to mesoderm, are mainly present in connective tissues and organ interstitium, and comprise: bone marrow, umbilical cord, fat, amnion, dental pulp, placenta, etc. Under proper conditions, the cell can be differentiated into fat, bone, cartilage and other tissue cells. Compared with MSC (mesenchymal stem cell) from bone marrow, the mesenchymal stem cells from umbilical cord, amnion and placenta have the advantages of large number of obtained cells, strong proliferation capacity, large immunoregulation effect, high total amount of secreted cell growth factors, convenience for amplification and passage, no problems of matching, rejection and the like due to convenient material acquisition and no moral and ethical dispute, are extremely suitable for clinical research and application, and are ideal sources of the mesenchymal stem cells.
However, mesenchymal stem cells need to be prepared, separated, efficiently purified and rapidly amplified in a GMP laboratory through a sample, and then cells meeting the standard can be produced. Therefore, the safe preservation of the sample becomes a basic requirement for the production of mesenchymal stem cells. From hospital delivery rooms to GMP laboratories, samples take hours as few as several days, and many days as many as several hours, which causes the number of cells in the samples to be small and the viability to be low due to the influence of factors such as time, temperature, nutrition, etc.
Therefore, the search for a universal preservation solution capable of preserving umbilical cord, amnion and placenta for a long time has been a problem to be solved by those skilled in the art.
Disclosure of Invention
In view of the above, the invention provides a universal preservation solution for human umbilical cord, amnion and placenta samples and a preparation method thereof.
The specific technical scheme of the invention is as follows:
a universal preservation solution for human umbilical cord, amnion and placenta samples comprises a DMEM basic culture medium, insulin, human serum albumin, vitamin C, EGF, penicillin, streptomycin and heparin sodium;
the pH value of the universal preservation solution is 7.5-7.8.
Preferably, the concentration of the insulin in the universal preservation solution is 10-20 mg/L;
the concentration of the human serum albumin in the universal preservation solution is 10-15 g/L;
the concentration of the vitamin C in the universal preservation solution is 100-200 mg/L;
the concentration of the EGF in the universal preservation solution is 10-50 mu g/L.
Preferably, the concentration of the insulin in the universal preservation solution is 10 mg/L;
the concentration of the human serum albumin in the universal preservation solution is 10 g/L;
the concentration of the vitamin C in the universal preservation solution is 100 mg/L;
the concentration of the EGF in the universal preservation solution is 10 mug/L.
Preferably, the concentration of the penicillin in the universal preservation solution is 50000-100000U/L;
the concentration of the streptomycin in the universal preservation solution is 50-100 mg/L;
the concentration of the heparin sodium in the universal preservation solution is 6250-12500U/L.
Preferably, the concentration of the penicillin in the universal preservation solution is 100000U/L;
the concentration of the streptomycin in the universal preservation solution is 100 mg/L;
the concentration of the heparin sodium in the universal preservation solution is 6250U/L.
The invention also provides a preparation method of the general preserving fluid in the technical scheme, which comprises the following steps:
adding human serum albumin and heparin sodium into a DMEM basic culture medium, sequentially and respectively adding penicillin, streptomycin, insulin, vitamin C and EGF, and filtering by adopting a filter membrane to obtain the universal preservation solution.
The invention also provides a method for preserving samples, which comprises the following steps:
the universal preservation solution is adopted to preserve the sample;
the sample is human umbilical cord, amnion and/or placenta.
Preferably, the cells of interest of the sample are mesenchymal stem cells.
Preferably, the storage time is 72 hours or less.
In conclusion, the invention provides a universal preservation solution which is composed of a DMEM basic culture medium, insulin, human serum albumin, vitamin C, EGF, penicillin, streptomycin and heparin sodium; the pH value of the universal preservation solution is 7.5-7.8. In the invention, the DMEM basic culture medium provides the most basic nutrition for the mesenchymal stem cells of the sample; the insulin can promote the mesenchymal stem cells of the sample to absorb and utilize sugar in the universal preservation solution and maintain the activity of the cells; the human serum albumin provides dynamic balance for the mesenchymal stem cells and maintains the osmotic pressure of the cells; the vitamin C can prevent the damage of free radicals generated by long-time oxidation to the mesenchymal stem cells; EGF can promote the proliferation of mesenchymal stem cells and maintain the original form of the cells; penicillin and streptomycin can prevent sample pollution caused by bacterial breeding; the heparin sodium can prevent blood from coagulating into blood clots and reduce metabolism; the pH value of the universal preservation solution is 7.5-7.8, and a proper acid-base environment is provided for the mesenchymal stem cells. Experimental results show that the universal preservation solution can be used for preserving umbilical cords, amnions and placentas for a long time, the obtained mesenchymal stem cells are large in number and high in survival rate, and human serum albumin, insulin, vitamin C and EGF have a synergistic effect in improving the number of the mesenchymal stem cells in the umbilical cords to be preserved and the survival rate of the mesenchymal stem cells. In addition, the quality guarantee time of the universal preservation solution can reach half a year, and the universal preservation solution can be prepared in a large amount at one time, so that the cost is controlled.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 is a diagram showing the viability of mesenchymal stem cells after umbilical cords are preserved for 24h, 48h and 72h by different preservation methods in example 2 of the present invention.
Detailed Description
The invention provides a universal preservation solution for human umbilical cord, amnion and placenta samples and a preparation method thereof.
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
Example 1
This example carried out the preparation of a universal preservative solution comprising the following steps:
adding 50mL of 20% human serum albumin (the concentration of the human serum albumin is 10g/50mL) and 1mL of heparin sodium injection into 950mLDMEM basal medium, uniformly mixing, sequentially adding 100000U of penicillin, 100mg of streptomycin, 10mg of insulin, 100mg of vitamin C and 10 mu g of EGF respectively, stirring for dissolving, filtering by using a 0.22 mu m sterile filter membrane to remove bacteria to obtain a general storage solution, marking the production date, and storing at 4 ℃.
Wherein, the concentration of the heparin sodium injection is 6250U/mL.
Comparative example 1
This comparative example was carried out to prepare a comparative universal preservation solution 1, prepared as in example 1, except that 50mL of 20% human serum albumin was replaced with an equal volume of PBS in the comparative universal preservation solution 1.
Comparative example 2
This comparative example was carried out to prepare a control universal stock solution 2 in the same manner as in example 1, except that the insulin was replaced with the same mass of PBS as that in the control universal stock solution 2.
Comparative example 3
This comparative example was carried out to prepare a comparative universal preservative solution 3 in the same manner as in example 1, except that the comparative universal preservative solution 3 was prepared by replacing vitamin C with PBS of equal mass.
Comparative example 4
This comparative example was carried out to prepare a comparative universal preservation solution 4 in the same manner as in example 1 except that the comparative universal preservation solution 4 was prepared by replacing EGF with PBS of equal mass.
Example 2
Under aseptic conditions, 24cm of newborn umbilical cords without infectious diseases and healthy in cesarean section operations are evenly divided into 8 parts, the parts are respectively stored in the universal storage solution, the normal saline, the DMEM culture medium, an aseptic container, the contrast universal storage solution 1, the contrast universal storage solution 2, the contrast universal storage solution 3 and the contrast universal storage solution 4 in example 1, umbilical cord tissues with the same length are taken after being stored at constant temperature of 4 ℃ for 24 hours, 48 hours and 72 hours, umbilical cord mesenchymal stem cells are separated by collagenase digestion, filtered and centrifuged, trypan is used for counting, and the cell viability is calculated.
Referring to fig. 1, table 1 and table 2, fig. 1 is a diagram of the viability of the mesenchymal stem cells of the umbilical cord after 24h, 48h and 72h are preserved by different preservation methods in example 2 of the present invention, table 1 is the number of the mesenchymal stem cells of the umbilical cord after 24h, 48h and 72h are preserved by different preservation methods, and table 2 is the viability of the mesenchymal stem cells of the umbilical cord after 24h, 48h and 72h are preserved by different preservation methods. The result shows that the umbilical cord can be preserved by the universal preservation solution in the embodiment 1 within 72h, the survival rate of the separated mesenchymal stem cells can be ensured to be more than 90% within 72h, the high activity of the umbilical cord mesenchymal stem cells can be maintained to the maximum extent, the extraction degree of the umbilical cord mesenchymal stem cells is improved, and the later-stage culture and cost saving of the mesenchymal stem cells are greatly improved. The physiological saline and the DMEM basic culture medium can preserve the umbilical cord within 24 hours, but the number and the survival rate of the obtained umbilical cord mesenchymal stem cells are gradually reduced along with the prolonging of time; the umbilical cord stored in the sterile container is lack of water in cells due to loss of liquid environment, so that the umbilical cord mesenchymal stem cells are rapidly apoptotic, and the living umbilical cord mesenchymal stem cells cannot be separated at 48 h. Compared with the control general-purpose preservation solution 1, the control general-purpose preservation solution 2, the control general-purpose preservation solution 3 and the control general-purpose preservation solution 4, the number of umbilical cord mesenchymal stem cells and the survival rate of the mesenchymal stem cells preserved by the general-purpose preservation solution in example 1 are obviously higher, which indicates that human albumin, insulin, vitamin C and EGF have synergistic effect in improving the number of umbilical cord mesenchymal stem cells and the survival rate of the mesenchymal stem cells.
TABLE 1 mesenchymal stem cell count after 24h, 48h, 72h preservation of umbilical cord using different preservation methods
| | 24h | 48h | 72h | ||
| Example 1 general-purpose preservative fluid | 7.17×106 | 6.78×106 | 6.36×106 | ||
| Physiological saline | 8.02×105 | 7.88×105 | 5.65×105 | ||
| DMEM medium | 4.32×106 | 3.01×106 | 1.94×106 | ||
| Sterile container | 3.86×103 | 0 | 0 | ||
| Reference universal preservative fluid 1 | 4.77×106 | 4.02×106 | 3.29×106 | ||
| Comparison general-purpose preservation solution 2 | 4.02×106 | 3.76×106 | 3.17×106 | ||
| Comparative general-purpose preservative fluid 3 | 4.57×106 | 2.99×106 | 1.88×106 | ||
| Comparison general-purpose preservation solution 4 | 4.98×106 | 4.19×106 | 3.67×106 |
TABLE 2 mesenchymal stem cell viability after 24h, 48h, 72h preservation of umbilical cord using different preservation methods
Example 3
One example of a healthy and infectious disease-free neonatal placenta obtained by a cesarean section was taken under aseptic conditions, the placental amniotic membrane was peeled off and evenly divided into 8 parts, which were stored in the universal preservation solution, physiological saline, a DMEM medium, an aseptic container, the control universal preservation solution 1, the control universal preservation solution 2, the control universal preservation solution 3 and the control universal preservation solution 4 of example 1, respectively, and the same mass of amniotic tissue was taken after being preserved at a constant temperature of 4 ℃ for 24 hours, 48 hours and 72 hours, and then the mesenchymal stem cells of the amniotic membrane were isolated by collagenase digestion, filtered, centrifuged, and counted using trypan blue.
The results refer to table 3, which shows that the universal preservation solution in example 1 has a good preservation effect on amnion, a large amount of mesenchymal stem cells can be separated in 72 hours, and the number of amnion in physiological saline and a DMEM basic culture medium is reduced along with the time extension; because the amnion contains a large amount of water, the amnion can still be preserved for 72h in a sterile container, but the number of mesenchymal stem cells is greatly reduced. Compared with the control universal preservation solution 1, the control universal preservation solution 2, the control universal preservation solution 3 and the control universal preservation solution 4, the number of the amnion mesenchymal stem cells preserved by the universal preservation solution in the embodiment 1 is obviously larger, which indicates that the human albumin, the insulin, the vitamin C and the EGF have a synergistic effect in increasing the number of the preserved amnion mesenchymal stem cells.
TABLE 3 mesenchymal stem cell count of amnion preserved for 24h, 48h, 72h by different preservation methods
| | 24h | 48h | 72h | ||
| Example 1 general-purpose preservative fluid | 6.66×106 | 6.17×106 | 5.83×106 | ||
| Physiological saline | 4.12×106 | 1.59×106 | 7.98×105 | ||
| DMEM medium | 5.52×106 | 3.07×106 | 1.50×106 | ||
| Sterile container | 3.23×105 | 1.14×104 | 3.04×103 | ||
| Reference universal preservative fluid 1 | 5.12×106 | 4.73×106 | 2.01×106 | ||
| Comparison general-purpose preservation solution 2 | 4.98×106 | 3.33×106 | 1.45×106 | ||
| Comparison general-purpose preservative fluid 3 | 5.56×106 | 4.90×106 | 1.99×106 | ||
| Comparison general-purpose preservation solution 4 | 5.88×106 | 4.91×106 | 2.49×106 |
Example 4
One example of a healthy and infectious disease-free placenta of a newborn subjected to a cesarean section operation was collected under aseptic conditions, the placenta was evenly divided into 8 portions, the portions were stored in the universal storage solution, physiological saline, a DMEM medium, an aseptic container, the control universal storage solution 1, the control universal storage solution 2, the control universal storage solution 3 and the control universal storage solution 4 of example 1, respectively, the placenta tissues of the same mass were collected after being stored at a constant temperature of 4 ℃ for 24 hours, 48 hours and 72 hours, the mesenchymal stem cells of the placenta were separated by collagenase digestion, filtered and centrifuged, and the cells were counted using trypan blue.
The results refer to table 4, which shows that the universal preservation solution of example 1 has a good preservation effect on placenta, can separate a large amount of mesenchymal stem cells within 72 hours, and the placenta in normal saline, a DMEM basic culture medium and a sterile container has large loss of mesenchymal stem cells. Compared with the control universal preservation solution 1, the control universal preservation solution 2, the control universal preservation solution 3 and the control universal preservation solution 4, the number of the mesenchymal stem cells in the placenta preserved by the universal preservation solution in the embodiment 1 is obviously larger, which indicates that the human serum albumin, the insulin, the vitamin C and the EGF have a synergistic effect in increasing the number of the mesenchymal stem cells in the placenta preserved.
TABLE 4 mesenchymal stem cell count after storing placenta for 24h, 48h, 72h by different storing methods
| | 24h | 48h | 72h | ||
| Example 1 general-purpose preservative fluid | 9.88×106 | 9.12×106 | 8.87×106 | ||
| Physiological saline | 5.98×105 | 1.26×105 | 8.44×104 | ||
| DMEM medium | 7.32×106 | 4.01×106 | 2.45×106 | ||
| Sterile container | 4.22×105 | 8.17×104 | 2.01×103 | ||
| Reference universal preservative fluid 1 | 7.08×106 | 5.07×106 | 3.81×106 | ||
| Comparison general-purpose preservative fluid 2 | 6.89×106 | 4.15×106 | 2.74×106 | ||
| Comparison general-purpose preservative fluid 3 | 7.88×106 | 5.48×106 | 3.91×106 | ||
| Comparison general-purpose preservation solution 4 | 7.46×106 | 4.22×106 | 2.31×106 |
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. The universal preservation solution for the human umbilical cord, amnion and placenta samples is characterized by comprising a DMEM basic culture medium, insulin, human serum albumin, vitamin C, EGF, penicillin, streptomycin and heparin sodium;
the pH value of the universal preservation solution is 7.5-7.8; the concentration of the insulin in the universal preservation solution is 10-20 mg/L;
the concentration of the human serum albumin in the universal preservation solution is 10-15 g/L;
the concentration of the vitamin C in the universal preservation solution is 100-200 mg/L;
the concentration of the EGF in the universal preservation solution is 10-50 mu g/L.
2. The universal preservation solution according to claim 1, wherein the concentration of insulin in the universal preservation solution is 10 mg/L;
the concentration of the human serum albumin in the universal preservation solution is 10 g/L;
the concentration of the vitamin C in the universal preservation solution is 100 mg/L;
the concentration of the EGF in the universal preservation solution is 10 mug/L.
3. The universal preservation solution according to claim 1, wherein the concentration of the penicillin in the universal preservation solution is 50000-100000U/L;
the concentration of the streptomycin in the universal preservation solution is 50-100 mg/L;
the concentration of the heparin sodium in the universal preservation solution is 6250-12500U/L.
4. The universal preservation solution according to claim 3, wherein the concentration of said penicillin in said universal preservation solution is 100000U/L;
the concentration of the streptomycin in the universal preservation solution is 100 mg/L;
the concentration of the heparin sodium in the universal preservation solution is 6250U/L.
5. The method for preparing a universal preservation solution for human umbilical cord, amniotic membrane and placental sample according to any one of claims 1 to 4, comprising the steps of:
adding human serum albumin and heparin sodium into a DMEM basic culture medium, sequentially and respectively adding penicillin, streptomycin, insulin, vitamin C and EGF, and filtering by adopting a filter membrane to obtain the universal preservation solution.
6. A method of preserving a sample, comprising the steps of:
preserving the sample by using the universal preserving fluid of any one of claims 1 to 4;
the sample is human umbilical cord, amnion and/or placenta.
7. The method of claim 6, wherein the cells of interest of the sample are mesenchymal stem cells.
8. The method of claim 6, wherein the holding time is 72 hours or less.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011438054.8A CN112471138B (en) | 2020-12-10 | 2020-12-10 | Universal preservation solution for human umbilical cord, amnion and placenta samples and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011438054.8A CN112471138B (en) | 2020-12-10 | 2020-12-10 | Universal preservation solution for human umbilical cord, amnion and placenta samples and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112471138A CN112471138A (en) | 2021-03-12 |
| CN112471138B true CN112471138B (en) | 2022-06-10 |
Family
ID=74941234
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011438054.8A Active CN112471138B (en) | 2020-12-10 | 2020-12-10 | Universal preservation solution for human umbilical cord, amnion and placenta samples and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112471138B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022203022A1 (en) * | 2021-03-26 | 2022-09-29 | 株式会社カネカ | Method for producing cell mass containing amnion-derived mesenchymal stem cells |
| CN113133444B (en) * | 2021-03-31 | 2022-10-11 | 北京益华生物科技有限公司 | Umbilical cord tissue cryopreservation liquid and preparation method thereof, umbilical cord tissue cryopreservation method and culture method |
| CN113287603B (en) * | 2021-06-18 | 2022-07-01 | 吉林省拓华生物科技有限公司 | Biological sample preservation solution and preparation method and application thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104212761A (en) * | 2014-08-20 | 2014-12-17 | 北京瑞思德生物科技有限公司 | Kit for preparing placenta stem cells |
| CN104542578A (en) * | 2015-02-05 | 2015-04-29 | 广州赛莱拉干细胞科技股份有限公司 | Cell preservation solution and preparation method and applications thereof |
| CN105779381A (en) * | 2016-03-30 | 2016-07-20 | 王凌仪 | Clinical treatment grade preparation method used for screening human umbilical cord derived WJ-MSCs (Wharton's jelly mesenchymal stem cells) in large scale by applying extracellular matrices through three-dimensional attachment and for cell treatment |
| CN106386782A (en) * | 2016-06-24 | 2017-02-15 | 安徽未名细胞治疗有限公司 | An umbilical cord preserving fluid |
| CN109423473A (en) * | 2017-08-21 | 2019-03-05 | 青岛瑞思德生物科技有限公司 | It is used to prepare the kit of placenta stem-cell |
| CN109479871A (en) * | 2017-09-12 | 2019-03-19 | 深圳华云生物科技发展有限公司 | Frozen stock solution, preparation method, application and China's Tong Shi glue tissue cryopreservation methods |
| CN109568344A (en) * | 2019-01-22 | 2019-04-05 | 药鼎(北京)国际细胞医学技术有限公司 | A kind of mesenchyme stem cell preserving fluid, preparation method and its application |
| CN110338187A (en) * | 2018-04-08 | 2019-10-18 | 生物角（厦门）科技有限公司 | A kind of mesenchymal stem cell serum-free frozen stock solution composition |
| CN110622956A (en) * | 2019-09-27 | 2019-12-31 | 广州南医大生物工程有限公司 | Umbilical cord mesenchymal stem cell preservation solution |
| CN112021304A (en) * | 2020-09-17 | 2020-12-04 | 山东省齐鲁干细胞工程有限公司 | Cryopreservation method and recovery method of umbilical cord mesenchymal stem cells |
-
2020
- 2020-12-10 CN CN202011438054.8A patent/CN112471138B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104212761A (en) * | 2014-08-20 | 2014-12-17 | 北京瑞思德生物科技有限公司 | Kit for preparing placenta stem cells |
| CN104542578A (en) * | 2015-02-05 | 2015-04-29 | 广州赛莱拉干细胞科技股份有限公司 | Cell preservation solution and preparation method and applications thereof |
| CN105779381A (en) * | 2016-03-30 | 2016-07-20 | 王凌仪 | Clinical treatment grade preparation method used for screening human umbilical cord derived WJ-MSCs (Wharton's jelly mesenchymal stem cells) in large scale by applying extracellular matrices through three-dimensional attachment and for cell treatment |
| CN106386782A (en) * | 2016-06-24 | 2017-02-15 | 安徽未名细胞治疗有限公司 | An umbilical cord preserving fluid |
| CN109423473A (en) * | 2017-08-21 | 2019-03-05 | 青岛瑞思德生物科技有限公司 | It is used to prepare the kit of placenta stem-cell |
| CN109479871A (en) * | 2017-09-12 | 2019-03-19 | 深圳华云生物科技发展有限公司 | Frozen stock solution, preparation method, application and China's Tong Shi glue tissue cryopreservation methods |
| CN110338187A (en) * | 2018-04-08 | 2019-10-18 | 生物角（厦门）科技有限公司 | A kind of mesenchymal stem cell serum-free frozen stock solution composition |
| CN109568344A (en) * | 2019-01-22 | 2019-04-05 | 药鼎(北京)国际细胞医学技术有限公司 | A kind of mesenchyme stem cell preserving fluid, preparation method and its application |
| CN110622956A (en) * | 2019-09-27 | 2019-12-31 | 广州南医大生物工程有限公司 | Umbilical cord mesenchymal stem cell preservation solution |
| CN112021304A (en) * | 2020-09-17 | 2020-12-04 | 山东省齐鲁干细胞工程有限公司 | Cryopreservation method and recovery method of umbilical cord mesenchymal stem cells |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112471138A (en) | 2021-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112471138B (en) | Universal preservation solution for human umbilical cord, amnion and placenta samples and preparation method thereof | |
| KR101514078B1 (en) | Methods for collecting and using placenta cord blood stem cells | |
| US9944900B2 (en) | Pulsatile perfusion extraction method for non-embryonic pluripotent stem cells | |
| WO2015025810A1 (en) | Production method and cryopreservation method for amniotic mesenchymal cell composition, and therapeutic agent | |
| JP5775667B2 (en) | Methods of lithium stimulation of cord blood stem cell proliferation and growth factor production | |
| CN107034183A (en) | The method for preparing autologous fat stem cell bioactivity peptide freeze-dried powder | |
| CN103667187A (en) | Isolated culture method of human adipose-derived stem cells and construction method of stem cell bank | |
| TW201510223A (en) | Method for separating living cell and constructing cell bank by means of tissue homogenate method | |
| CN112841168A (en) | Tissue preservation solution and application thereof | |
| CN111602652A (en) | Umbilical cord mesenchymal stem cell cryopreservation protective solution | |
| CN106614524B (en) | Preservation solution and preservation method for mesenchymal stem cells | |
| CN113287603B (en) | Biological sample preservation solution and preparation method and application thereof | |
| CN112195151B (en) | Dental pulp mesenchymal stem cell recovery culture solution, preparation method and recovery culture method | |
| CN109362709A (en) | A kind of placenta tissue saves liquid and preparation method thereof | |
| CN110839614B (en) | Autologous umbilical cord mesenchymal stem cell cryopreservation liquid and preparation and cryopreservation methods thereof | |
| CN111808807A (en) | Mesenchymal stem cell serum-free medium without need of pre-coating and application thereof | |
| CN107090434B (en) | Blood anticoagulation and preservation method for improving CIK cell culture effect | |
| CN108849857A (en) | A kind of transport protection liquid of umbilical cord mesenchymal stem cells | |
| CN111575229B (en) | Separation method of placenta decidua stem cells | |
| CN114557336B (en) | Tissue treatment fluid capable of improving primary separation quantity and activity of tissues as well as preparation method and application thereof | |
| CN116083340A (en) | Preparation method of silkworm haemolymph liquid for inhibiting blackening reaction and reducing endotoxin | |
| US20190357524A1 (en) | Preservative solution for human stem cells, human stem cell suspension, and method for preserving human stem cells | |
| US20240301354A1 (en) | System and method for converting adipose derived mesenchymal stems cells to hematopoietic stem/progenitor cells and differentiating into blood cells and applications of same | |
| WO2022203022A1 (en) | Method for producing cell mass containing amnion-derived mesenchymal stem cells | |
| CN114097768A (en) | Protective solution for bone marrow stem cells and preparation method and application thereof |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230925 Address after: Room 205, 2nd Floor, Building 21, No. 333 Nanshan Road, Jianghai District, Jiangmen City, Guangdong Province, 529000 Patentee after: Guangdong Factor Life Medical Technology Co.,Ltd. Address before: Room 202 on the first and second floors of Building 21 (2 # comprehensive supporting buildings), Industrial Acceleration Park, No. 333 Nanshan Road, Jianghai District, Jiangmen City, Guangdong Province, 529000 (information declaration system) Patentee before: Dude (Jiangmen) Biotechnology Co.,Ltd. |
|
| TR01 | Transfer of patent right |