CN114457011A - Stem cell culture medium and stem cell separation culture method - Google Patents
Stem cell culture medium and stem cell separation culture method Download PDFInfo
- Publication number
- CN114457011A CN114457011A CN202210170433.6A CN202210170433A CN114457011A CN 114457011 A CN114457011 A CN 114457011A CN 202210170433 A CN202210170433 A CN 202210170433A CN 114457011 A CN114457011 A CN 114457011A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- cells
- culture medium
- culture
- medium
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/38—Vitamins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/80—Undefined extracts from animals
- C12N2500/84—Undefined extracts from animals from mammals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Abstract
The invention discloses a stem cell culture medium and a stem cell separation culture method, wherein the stem cell culture medium comprises the following components: 45-50% of basal medium DMEM, 5-10% of fetal bovine serum, 20-25% of Baitong stem cell culture medium, 20-25% of alpha-MEM, 50ng/mL of bFGF, 50ng/mL of EGF and 10mg/mL of VC. The stem cell culture medium is prepared by mixing various culture media according to a certain proportion, and the activity and purity of cultured cells reach more than 95 percent on the basis of lower cost; a larger total cell amount can be achieved at a lower generation number; the culture medium has good culture effect on stem cells from various sources, and particularly has remarkable effect in the culture of umbilical cord mesenchymal stem cells from various sources; by combining the corresponding stem cell separation culture method, the total number of 100 hundred million cells can be reached in the fourth generation by averaging one umbilical cord.
Description
Technical Field
The invention relates to the technical field of stem cells, in particular to a stem cell culture medium and a stem cell separation culture method.
Background
The stem cell is a cell with multiple differentiation potentials, and can be widely applied to the fields of pharmacology, tissue and organ repair and cell therapy. The main sources of stem cells are umbilical cord, embryo, bone marrow, fat, dental pulp, multipotentiality and self-renewal are the basic characteristics of stem cells. Specifically, stem cells have the following biological characteristics:
1. belongs to non-terminally differentiated cells, maintains undifferentiated or poorly differentiated characteristics throughout life, and lacks differentiation markers. 2. The number of positions on the body is relatively constant. 3. Has self-renewal capability. 4. Can divide and proliferate indefinitely, can be in a static state for a long time, and stem cells can continuously divide for several generations. 5. Has multidirectional differentiation potential and can be differentiated into various tissue cells of different types; also has the plasticity of differentiation and development, and under specific circumstances can be induced to differentiate into developmentally unrelated cell types, the differentiation of which is influenced by the stem cell niche of the surrounding microenvironment. 6. Slow periodicity of the fragmentation. 7. The stem cells grow in two ways, one is symmetrical division to form two identical stem cells, the other is asymmetrical division, one of the asymmetrical division maintains the characteristics of parents and still remains as stem cells, and the other daughter cells irreversibly trend to the terminal of differentiation to become function-specific differentiated cells.
Stem cells have the following functions:
1. replacement and repair of dead and damaged cells
The cells have the characteristic of automatic homing, and can be gathered to damaged organs and relevant parts and differentiated into specific cells after being injected into a human body. After the stem cells are injected into a human body, new cells and tissues grow in the microenvironment of the target tissues, and damaged tissues are repaired. The stem cell self-differentiation function is utilized to grow new tissue cells, so that the aging, death and damage of the tissue cells are compensated, and diseased tissues and cells are recovered to be normal. Adding new blood vessel to form microcirculation and improve injured tissue.
2. Paracrine action
After stem cells are injected into tissues, various bioactive factors such as various factors including neurotrophic factors and anti-apoptosis factors, regulatory peptides, gas signal molecules and the like can be produced, and the stem cells can act on peripheral cells. Can promote cell proliferation, inhibit apoptosis of functional cells, differentiate the existing tissue progenitor cells into tissue cells, repair damaged tissues and grow new tissues.
3. Activating dormant and suppressed cells
Growth and development of the body is accomplished by cell division, when cells divide, a portion of which breaks away from the normal cell cycle and represents functional sleep. Some cells are affected by intrinsic factors and growth is inhibited. The cells in the dormant or inhibited state can not divide and proliferate any more, but only enter the aging metabolic process, so that the new cells of the human body are reduced, the metabolism is slowed down, and the human body enters the aging stage. With age, stem cells transmit youthful signals to adult cells that escape the cell cycle. After dormancy and activation of the inhibition cells stimulated by stem cells, the cells enter the cell cycle, and increase the number of new cells in the body through division and proliferation, so that the metabolic process of the human body is recovered to normal or even reversed.
The umbilical cord mesenchymal stem cells have wide sources, easily obtained materials, convenient transplantation and strong proliferation capacity, can always keep the multidirectional differentiation potential in the in vitro long-term culture process, can be differentiated into osteogenesis, cartilage, tendon, muscle cells, fat cells, nerve cells, liver cells and the like under the induction of specific conditions, and are ideal tissue engineering seed cells.
At present, culture media for stem cells in the market are expensive, the culture media are consumed for the operation and research of a large amount of cultured stem cells, the culture media prepared by using basic culture media in most laboratories are cheap, but the culture effect is poor, the state of the cultured cells is poor after multiple passages, and the total number of the cells is low.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a stem cell culture medium and a stem cell separation culture method, which can solve the technical problems.
(II) technical scheme
In order to solve the above technical problems, the present invention provides the following technical solutions: a stem cell culture medium comprising the following components: 45-50% of basal medium DMEM, 5-10% of fetal bovine serum, 20-25% of Baitong stem cell culture medium, 20-25% of alpha-MEM, 50ng/mL of bFGF, 50ng/mL of EGF and 10mg/mL of VC.
Preferably, the stem cell culture medium comprises the following components: 45% by volume of a basal medium DMEM, 5% by volume of fetal bovine serum, 25% by volume of a Baitong stem cell medium, 25% by volume of an alpha-MEM medium, 50ng/mL of bFGF, 50ng/mL of EGF and 10mg/mL of VC.
Preferably, the basal medium DMEM is a low-sugar type.
In order to solve the above technical problem, the present invention provides another technical solution as follows: a stem cell separation culture method comprises the following steps:
s1: cleaning the umbilical cord with 100ml of 0.9% normal saline, pouring off the normal saline, completely soaking the umbilical cord with 100ml of 75% alcohol for 3-5 minutes, pouring off the alcohol, cleaning off residual alcohol with 100ml of 0.9% normal saline, and pouring off the normal saline;
s2: cutting umbilical cord into 3cm pieces, peeling two arteries and a vein in each piece with forceps, removing epidermis, retaining jelly-like substance between artery and vein, cutting into 3 × 3mm tissue blocks with scissors, placing in 9cm culture dish, adhering wall for 1 hr, adding 10ml of the dry cell culture medium of any one of claims 1-3, standing at 37 deg.C and 5% CO2The incubator for 5-6 days;
s3: after the static culture is completed for 5-6 days in step S2, the old stem cell culture medium is poured out of the culture dish, the first liquid change is performed and the new stem cell culture medium is poured out, and the culture dish is placed at 37 ℃ again and 5% CO2The incubator is used for culturing;
s4: after 3-4 days of observation at step S3, old stem cells in the culture dish were poured outThe culture medium is changed for the second time, new stem cell culture medium is poured into the culture dish, and the culture dish is placed at 37 ℃ and 5% CO2The incubator is used for culturing;
s5: when the cell density in the culture dish reaches 50%, the tissue piece in the culture dish is transferred to a new culture dish with tweezers, and the cells in the old culture dish are subjected to the passaging operation, which is observed after 3-4 days in step S4: washing with PBS buffer solution, digesting with 0.25% pancreatin for 2min, adding stem cell culture medium to terminate reaction, transferring cell suspension into centrifuge tube, centrifuging, removing supernatant, adding stem cell culture medium to dilute cells to 1 × 106Adding into culture flask, further standing at 37 deg.C and 5% CO2The incubator is used for culturing;
s6: a passaging operation of the cells in the new culture dish in S5 when the cell density in the new culture dish in S5 reaches 50% after 2 to 3 days in step S5; and meanwhile, observing the condition of the cells passaged to the culture bottle in the S5, and when the cell density reaches more than 80%, carrying out secondary passage operation on the cells passaged to the culture bottle or carrying out cryopreservation operation on the cells serving as seed cells.
(III) advantageous effects
Compared with the prior art, the invention provides a stem cell culture medium and a stem cell isolated culture method, which have the following beneficial effects: the stem cell culture medium is prepared by mixing various culture media according to a certain proportion, and the activity and purity of cultured cells reach more than 95 percent on the basis of lower cost; the subculture expansion ratio of the general stem cells is 1: 3, the average amplification ratio of the stem cell culture medium of the invention is 1: 6, the larger total amount of cells can be reached at a lower generation number, and the requirements of various subsequent experiments are met; the culture medium has good culture effect on stem cells from various sources, and particularly has remarkable effect in the culture of umbilical cord mesenchymal stem cells from various sources; by combining the corresponding stem cell separation culture method, the total number of 100 hundred million cells can be reached in the fourth generation by averaging one umbilical cord.
Drawings
FIG. 1 is a graph comparing the activity of different media at each stage;
FIG. 2 is a microscopic image of P4 cells in different media (4X);
FIG. 3 is a photograph (10X) of microscopic observation of P4 cells in different media;
FIG. 4 is a photograph (20X) of microscopic observation of P4 cells in different media.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a stem cell culture medium, which comprises the following components: 45-50% of basal medium DMEM, 5-10% of fetal bovine serum, 20-25% of baitong stem cell culture medium, 20-25% of alpha-MEM, 50ng/mL of bFGF, 50ng/mL of EGF and 10mg/mL of VC.
Preferably, the stem cell culture medium comprises the following components: 45% by volume of a basal medium DMEM, 5% by volume of fetal bovine serum, 25% by volume of a Baitong stem cell medium, 25% by volume of an alpha-MEM medium, 50ng/mL of bFGF, 50ng/mL of EGF and 10mg/mL of VC.
The basal medium DMEM is a medium containing various amino acids and glucose, is developed on the basis of the MEM medium, increases the dosage of various components compared with the MEM, and is divided into a high-sugar type (lower than 4500mg/L) and a low-sugar type (lower than 1000 mg/L); preferably, the stem cell culture medium of the present invention uses a low-sugar type basal medium DMEM.
The fetal calf serum is a slightly viscous liquid with characteristics, light yellow and clear appearance, no hemolysis and no foreign matters, is taken from a fetal calf born by cesarean section, is a natural culture medium with the largest dosage in cell culture, contains rich nutrient components necessary for cell growth, is commonly used for in vitro culture of animal cells, and has extremely important functions.
The Baitong stem cell culture medium is a stem cell culture medium which is generally used in the market and is produced by Hangzhou Baitong biotechnology limited company; the alpha-MEM culture medium is a modified MEM culture medium, and the Baitong stem cell culture medium and the alpha-MEM culture medium can be purchased from the market.
bFGF is human basic fibroblast growth factor, EGF is human epidermal growth factor, and VC is vitamin C.
The invention also provides a stem cell separation culture method, which comprises the following steps:
s1: the umbilical cord is washed by 100ml of 0.9% normal saline, the normal saline is poured off, 100ml of 75% alcohol is used for completely soaking the umbilical cord for 3-5 minutes, the alcohol is poured off, 100ml of 0.9% normal saline is used for washing off residual alcohol, and the normal saline is poured off.
S2: cutting umbilical cord into 3cm long segments, peeling off two arteries and one vein in each segment with forceps, removing epidermis, retaining Jelly-like substance (Wharton's Jelly) between artery and vein, cutting into 3 × 3mm tissue blocks, placing in 9cm culture dish, adhering for 1 hr, adding 10ml of the dry cell culture medium of any one of claims 1-3, and standing at 37 deg.C and 5% CO2The incubator for 5-6 days; no observation or other operation is needed in the middle.
S3: after the static culture is completed for 5-6 days in step S2, observation is performed, the stem cells can be seen to grow sporadically under a microscope, the old stem cell culture medium in the culture dish is poured out, the first liquid change is performed, the new stem cell culture medium is poured in, the culture dish is placed at 37 ℃ again, and 5% CO is added2The incubator of (2) for cultivation.
S4: after 3-4 days at step S3, the density of cells in the dish is typically around 30%, and the old dry fine particles are removed from the dishCell culture medium, changing the culture medium for the second time, pouring new stem cell culture medium, and placing the culture dish at 37 deg.C and 5% CO2The incubator of (2) for cultivation.
S5: when the cell density in the culture dish reaches 50%, which is observed after 3-4 days in step S4, about 70% of the cell density in the culture dish can be generally seen at this time, the tissue mass in the culture dish is transferred to a new culture dish with tweezers, and the cells in the old culture dish are subcultured: washing with PBS buffer solution, digesting with 0.25% pancreatin for 2min, adding stem cell culture medium to terminate reaction, transferring cell suspension into centrifuge tube, centrifuging, removing supernatant, adding stem cell culture medium to dilute cells to 1 × 106The resulting solution was concentrated in a culture flask, and further placed at 37 ℃ with 5% CO2The incubator of (2) for cultivation.
S6: observing the cell adherence condition in the new culture dish 2-3 days after the step S5, and performing a first passage operation on the cells in the new culture dish in S5 when the cell density in the new culture dish in S5 reaches 50%, which is about 70% in general, and the operation can refer to the passage operation in S5; meanwhile, when the cell density of the cells passaged to the culture flask in S5 is observed to be 80% or more, the cell density of the culture flask is generally about 90%, and the cells passaged to the culture flask are subjected to a second passaging operation or a cryopreservation operation as seed cells.
To maintain good stem cell characteristics, stem cells are generally passed to the fourth passage and frozen in their entirety.
Compared with the prior art, the invention provides a stem cell culture medium and a stem cell isolated culture method, which have the following beneficial effects: the stem cell culture medium is prepared by mixing various culture media according to a certain proportion, and the activity and purity of cultured cells reach more than 95 percent on the basis of lower cost; the subculture expansion ratio of the general stem cells is 1: 3, the average amplification ratio of the stem cell culture medium of the invention is 1: 6, a larger total amount of cells can be achieved at a lower generation number, and the requirements of various subsequent experiments are met; the culture medium has good culture effect on stem cells from various sources, and particularly has remarkable effect in the culture of umbilical cord mesenchymal stem cells from various sources; by combining the corresponding stem cell separation culture method, the total number of 100 hundred million cells can be reached in the fourth generation by averaging one umbilical cord.
Statistics of the test results were performed for different media as follows, medium a: baitong stem cell medium (medium commonly used on the market), medium B: DMEM + 10% PBS +, bFGF 100ng/ml, EGF100ng/ml (laboratory used medium), medium C: the stem cell culture medium of the invention.
As can be seen from FIG. 1, medium C was more active than medium A, B at different times.
Regarding the differentiation-inducing ability of the stem cell culture medium of the present invention, adipogenic induction differentiation: after the cells are attached to 100% confluence, adding an interstitial stem cell adipogenesis induction liquid, after adipogenesis induction is carried out for 72 hours, small lipid droplets appear in the cells, the number of the lipid droplets is increased and the lipid droplets are fused with each other after about 2 weeks, the cells are changed into a circle or a polygon from a long fusiform, and oil red O staining shows that a large amount of lipid precipitates. Osteogenic induced differentiation: after the cells are attached to the wall and are about 70 percent confluent, adding mesenchymal stem cell osteogenesis inducing liquid, and after induced culture is carried out for 3 days, increasing the cell volume to form a short spindle shape; on induction day 7, the cells were polygonal and cytoplasmic with increased cell granules; 14 days, the cytoplasm is full of particles, the cells grow in colony shape, and calcium deposition can be seen among the cells; in 29 days, cells in the center of cell nodules gradually fuse and lose cell structures, calcium nodules are obviously formed, and the calcium nodules are stained by alizarin red to form red nodules.
It is to be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (4)
1. A stem cell culture medium comprising the following components: 45-50% of basal medium DMEM, 5-10% of fetal bovine serum, 20-25% of Baitong stem cell culture medium, 20-25% of alpha-MEM, 50ng/mL of bFGF, 50ng/mL of EGF and 10mg/mL of VC.
2. The stem cell culture medium of claim 1, wherein: the stem cell culture medium comprises the following components: the basic medium DMEM with the volume percentage of 45%, the fetal calf serum with the volume percentage of 5%, the Baitong stem cell medium with the volume percentage of 25%, the alpha-MEM medium with the volume percentage of 25%, the bFGF with the concentration of 50ng/mL, the EGF with the concentration of 50ng/mL and the VC with the concentration of 10 mg/mL.
3. The stem cell culture medium of claim 2, wherein: the basal medium DMEM is low-sugar type.
4. A stem cell isolation and culture method is characterized by comprising the following steps:
s1: cleaning the umbilical cord with 100ml of 0.9% normal saline, pouring the normal saline, completely soaking the umbilical cord with 100ml of 75% alcohol for 3-5 minutes, pouring the alcohol, cleaning residual alcohol with 100ml of 0.9% normal saline, and pouring the normal saline;
s2: cutting the umbilical cord into 3cm pieces, and then using forceps to cut each pieceStripping off two arteries and one vein in the section, removing epidermis, retaining gel-like substance between the arteries and veins, cutting into 3 × 3mm tissue blocks with scissors, placing in 9cm culture dish, adhering for 1 hr or so, adding 10ml of the dry cell culture medium of any one of claims 1-3, and standing at 37 deg.C and 5% CO25-6 days in the incubator;
s3: after the static culture for 5-6 days is completed in the step S2, the old stem cell culture medium in the culture dish is poured out, the first liquid change is carried out, the new stem cell culture medium is poured in, and the culture dish is placed at 37 ℃ again and 5% CO2The incubator is used for culturing;
s4: observing after 3-4 days of the step S3, pouring out the old stem cell culture medium in the culture dish, carrying out a second liquid change, pouring in a new stem cell culture medium, and putting the culture dish at 37 ℃ again and 5% CO2The incubator is used for culturing;
s5: when the cell density in the culture dish reaches 50%, the tissue piece in the culture dish is transferred to a new culture dish with forceps, and the cells in the old culture dish are subjected to subculture operation, which is observed after 3-4 days in the step S4: washing with PBS buffer solution, digesting with 0.25% pancreatin for 2min, adding the stem cell culture medium to terminate reaction, transferring the cell suspension to a centrifuge tube for centrifugation, pouring off the supernatant after centrifugation, adding the stem cell culture medium to dilute the cells into 1 × 106The resulting solution was concentrated in a culture flask, and further placed at 37 ℃ with 5% CO2The incubator is used for culturing;
s6: after 2-3 days of the step S5, when the cell density in the new culture dish of S5 reaches 50%, performing the passaging operation on the cells in the new culture dish of S5; and meanwhile, observing the condition of the cells passaged to the culture bottle in the S5, and when the cell density reaches more than 80%, carrying out the second passage operation on the cells passaged to the culture bottle or carrying out cryopreservation operation on the cells serving as seed cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210170433.6A CN114457011A (en) | 2022-02-24 | 2022-02-24 | Stem cell culture medium and stem cell separation culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210170433.6A CN114457011A (en) | 2022-02-24 | 2022-02-24 | Stem cell culture medium and stem cell separation culture method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114457011A true CN114457011A (en) | 2022-05-10 |
Family
ID=81415800
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210170433.6A Pending CN114457011A (en) | 2022-02-24 | 2022-02-24 | Stem cell culture medium and stem cell separation culture method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114457011A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101412985A (en) * | 2007-10-15 | 2009-04-22 | 华东理工大学 | Serum-free medium for in vitro cultivation and amplification of mesenchymal stem cells |
| CN104630142A (en) * | 2015-02-06 | 2015-05-20 | 广州赛莱拉干细胞科技股份有限公司 | Separation and culture method of bovine umbilical cord mesenchymal stem cells |
| CN106318979A (en) * | 2015-02-10 | 2017-01-11 | 黄兵 | Method for inducing transdifferentiation of mesenchymal stem cells into skin stem cells |
| CN107586757A (en) * | 2017-11-01 | 2018-01-16 | 天津昂赛细胞基因工程有限公司 | One boar Mesenchymal stem cell nutrient solution and preparation method thereof |
-
2022
- 2022-02-24 CN CN202210170433.6A patent/CN114457011A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101412985A (en) * | 2007-10-15 | 2009-04-22 | 华东理工大学 | Serum-free medium for in vitro cultivation and amplification of mesenchymal stem cells |
| CN104630142A (en) * | 2015-02-06 | 2015-05-20 | 广州赛莱拉干细胞科技股份有限公司 | Separation and culture method of bovine umbilical cord mesenchymal stem cells |
| CN106318979A (en) * | 2015-02-10 | 2017-01-11 | 黄兵 | Method for inducing transdifferentiation of mesenchymal stem cells into skin stem cells |
| CN107586757A (en) * | 2017-11-01 | 2018-01-16 | 天津昂赛细胞基因工程有限公司 | One boar Mesenchymal stem cell nutrient solution and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| BORZO GHARIBI ET AL.: ""Effects of Medium Supplements on Proliferation,Differentiation Potential, and In Vitro Expansion of Mesenchymal Stem Cells"", 《STEM CELLS TRANSLATIONAL MEDICINE》 * |
| 林田 等: ""人脐带间充质干细胞培养体系的优化"", 《广西医科大学学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101077760B1 (en) | Progenitor cells from wharton's jelly of human umbilical cord | |
| RU2306335C2 (en) | Stem cells and matrices obtained from adipose tissue | |
| JP5785394B2 (en) | Optimized and defined method for isolating and storing progenitor cells from human umbilical cord | |
| US20110217385A1 (en) | Method for extracting mesenchymal stem cell from human or animal embryo and for extracting the secretion product thereof | |
| KR20140000945A (en) | Method for preparing high concentration of stem cells | |
| WO2011101834A1 (en) | A method for obtaining mesenchymal stem cells, media, methods and composition thereof | |
| CN110898078B (en) | Preparation and application of mesenchymal stem cell secretion factor | |
| CN105779384A (en) | Seed cell screening and culturing cryopreservation technical method of human amniotic mesenchymal stem cells for tissue engineering | |
| WO2020130038A1 (en) | Composition for regenerative therapy and method of producing composition for regenerative therapy | |
| CN109652368B (en) | Method for obtaining primary mesenchymal stem cells from umbilical cord tissue | |
| WO2003050273A1 (en) | Human cell culture medium and culture method | |
| CN112063580B (en) | Culture inducing liquid for umbilical cord and placenta mesenchymal stem cells | |
| JP6721504B2 (en) | Process for producing pluripotent stem and progenitor cells | |
| Gao et al. | Putative epidermal stem cell convert into corneal epithelium-like cell under corneal tissue in vitro | |
| CN114457011A (en) | Stem cell culture medium and stem cell separation culture method | |
| CN115125192A (en) | Bone marrow supernatant and application thereof in cell culture | |
| CN106701666A (en) | Inducing liquid and method for inducing human amniotic mesenchymal stem cells to differentiate to cardiomyocyte-like cells | |
| KR20080094431A (en) | Method for differentiating, culturing and isolating neural progenitor cells from peripheral blood mononuclear cells | |
| US20090148416A1 (en) | Angiogenically induced transplants and methods for their use and manufacture | |
| KR102435452B1 (en) | Mesenchymal stem cells with excellent anti-senescence ability and stemness property, and culture method of the same | |
| KR102087084B1 (en) | Culture method of adipose-drived stem cell proliferation expressing anti-aging factors | |
| EP4134429A1 (en) | Stem cell culturing method for promoting initial yield of stem cel.ls | |
| Nurhayati et al. | Brief comparative study on the isolation and culture methods of human keratinocyte from skin tissue | |
| CN100519741C (en) | Extract of egg celsl from animals | |
| Suyoko et al. | Simple Procedure for the Isolation of Mesenchymal Stem Cells from Different Parts of the Human Umbilical Cord |
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 |