CN106318906A - Method for large-scale culture of human umbilical cord mesenchymal stem cells - Google Patents
Method for large-scale culture of human umbilical cord mesenchymal stem cells Download PDFInfo
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- CN106318906A CN106318906A CN201611051086.6A CN201611051086A CN106318906A CN 106318906 A CN106318906 A CN 106318906A CN 201611051086 A CN201611051086 A CN 201611051086A CN 106318906 A CN106318906 A CN 106318906A
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- 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
- C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
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- 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
- C12N2531/00—Microcarriers
Abstract
The invention relates to the field of stem cells and discloses a method for large-scale culture of human umbilical cord mesenchymal stem cells. The method includes: adding an umbilical cord mesenchymal stem cell culture medium into a glass culture bottle, and performing incubation balance at 35.5 DEG C for 30min in an incubator with 5% of CO2, wherein humidity of the incubation is 95%; adjusting the umbilical cord mesenchymal stem cell culture medium to be 35 DEG C in temperature and 7.05 in pH, adding pretreated microcarriers and umbilical cord mesenchymal stem cells, and stirring for 4h at 10RPM-40RPM; heating to 37 DEG C, culturing for 96h at 35RPM-60RPM under the condition of 5% of CO2, and replenishing the umbilical cord mesenchymal stem cell culture medium in the process. Compared with the prior art, the method for large-scale culture of the human umbilical cord mesenchymal stem cells has advantages that more cells are obtained after proliferation, cell activity and proliferation capacity are improved, and great stem cell characteristics and morphology of the umbilical cord mesenchymal stem cells can be well kept.
Description
Technical field
The present invention relates to stem cell field, a kind of method being specifically related to large-scale culture human umbilical cord mesenchymal stem cells.
Background technology
Mescenchymal stem cell (mesenchymal stem cells, MSCs) is derived from growing mesoderm in early days with outer
Germinal layer, has the cell of the characteristics such as self renewal, Multidirectional Differentiation, pluripotency, hematopoiesis support and immunoregulation, produces
Raw also releasing nutrients material, promotes cytothesis and angiogenesis by paracrine approach, has important in regenerative medicine field
Effect.
At present, from fat, bone marrow, umbilical cord, placenta tissue, mescenchymal stem cell is mainly extracted.Mescenchymal stem cell is continuous
Still there is multi-lineage potential, for the various disease of clinical treatment, such as nervous system disease, kidney after Secondary Culture and freezen protective
The treatment of disease, autoimmune disease, malignant tumor etc. provides new approach.
Umbilical cord mesenchymal stem cells (UCMSCs) is present in umbilical cord Wal and leads to glue (Wharton'sjelly) and blood vessel week
Enclose a kind of stem cell in tissue.Derive from the mescenchymal stem cell of umbilical cord, draw materials conveniently, abundance, it is easy to gather and transport
Defeated, biological characteristics is stable, and immunogenicity is low, without allosome rejection, and low cost, harmless to donor and be not related to ethics
The advantages such as problem so that it is become following stem cell and there is in medical applications the ideal chose of great potential.
But umbilical cord mesenchymal stem cells is either still required for substantial amounts of cell for organizational project for clinical.Face
Bed application often needs to reach 108-1010The order of magnitude.Owing to mescenchymal stem cell needs adherent growth, the cell quantity of this scale
If using culture bottle amplification in the lab, consumptive material consumption is big, and artificial paying also is surprising.
Bioreactor is the biological function utilizing organism to be had, in vitro or in vivo by biochemical reaction or biology
The metabolism of self obtains the apparatus system of target product, cell, histoorgan etc..Bioreactor can be used for expanding on a large scale
Cell, but generally bioreactor is for the extensive amplification of suspension cell, and be open system, need a lot of support
Equipment, not only operates complexity, is also easy to cause the problems such as pollution, clinical practice to there is sizable risk.
Summary of the invention
In view of this, the problem that present invention aim at existing for prior art, it is provided that a kind of large-scale culture people's umbilicus
The cultural method of the method umbilical cord mesenchymal stem cells with mescenchymal stem cell, the method for the invention can obtain the short time in a large number
High-purity umbilical cord mesenchymal stem cells, and the stem cell properties that umbilical cord mesenchymal stem cells is good can be kept.
In order to realize the purpose of the present invention, the present invention adopts the following technical scheme that
A kind of method of large-scale culture human umbilical cord mesenchymal stem cells, comprises the steps:
1) in glass culture bottle, umbilical cord mesenchymal stem cells culture medium is added, at humidity 95%, 5%CO2Incubator
In balance 30 minutes in 35.5 DEG C of incubations；
2) adjust umbilical cord mesenchymal stem cells culture medium temperature to 35 DEG C, pH7.05, add pretreatment microcarrier and
Umbilical cord mesenchymal stem cells, the speed stirring 4h of 10RPM～40RPM；
3) 37 DEG C it are warming up to, in mixing speed, the 5%CO of 35RPM～60RPM2Under the conditions of cultivate 96h, period adds umbilicus
Interband mesenchymal stem cell media.
In some embodiments, pre-described in the method for large-scale culture human umbilical cord mesenchymal stem cells of the present invention
The addition of the microcarrier processed is to the final concentration of 0.5mg/mL-5.0mg/mL of microcarrier.
In some embodiments, micro-described in the method for large-scale culture human umbilical cord mesenchymal stem cells of the present invention
Carrier is that Cytodex1 microcarrier, Cytodex2 microcarrier, Cytodex3 microcarrier, Cytopore microcarrier or Cytoline are micro-
Carrier.In some embodiments, the method for described microcarrier pretreatment is that microcarrier is first by the phosphoric acid buffer immersion of pH 7.4
Bubble expands, sterilizing；Before using, resuspended by a small amount of culture medium after the culture medium rinse of aseptic microcarrier warm.
In some embodiments, the method for described microcarrier pretreatment is that microcarrier is first with the phosphate buffer of pH 7.4
Soak and expand, sterilizing；Before using, resuspended by a small amount of culture medium after the culture medium rinse of aseptic microcarrier warm.
In certain embodiments, the method for described microcarrier pretreatment is that dry microcarrier is without Ca2+、Mg2+、
The phosphate buffer of pH 7.4 at room temperature soaks and expands at least 3 hours, abandoning supernatant, with new preparation without Ca2+、Mg2 +, the PBS of pH 7.4 wash microcarrier several minutes, discard PBS, change new without Ca2+、Mg2+, the PBS of pH 7.4, the most micro-load
Liquid solution autoclaving sterilizing；Before use, first allowing aseptic microcarrier settle, incline supernatant, then with the training of warm
Support base rinse, then allow microcarrier settle, supernatant discarded, then resuspended rear standby by a small amount of culture medium.
In some embodiments, umbilicus described in the method for large-scale culture human umbilical cord mesenchymal stem cells of the present invention
Addition with mescenchymal stem cell is to umbilical cord mesenchymal stem cells final concentration of 0.5 × 105cells/mL-2×
105cells/mL。
Preferably, step 2) described stirring carries out under lucifuge.
In the method for large-scale culture human umbilical cord mesenchymal stem cells of the present invention, need in the training period to add and fill between umbilical cord
Matter stem cell media.In some embodiments, add umbilical cord mesenchymal stem cells culture medium described in and be specially 48h replacing
The culture medium of 50%.
Umbilical cord mesenchymal stem cells culture medium described in the method for large-scale culture human umbilical cord mesenchymal stem cells of the present invention
Formula be DMEM/F12+15%FBS.
The acquisition methods of umbilical cord mesenchymal stem cells of the present invention is: the human umbilical cord mesenchymal stem cells of cultivation stops stirring
Mix, microcarrier settle, supernatant discarded culture medium, with pH7.6 containing EDTA without Ca2+、Mg2+The PBS washing of ion, then
Use pancreas enzyme-EDTA solution digestion, after 15min, add the cell culture fluid containing 10% (v/v) serum FBS and terminate digestion, obtain
P2, for cell, can get 1.6 × 106Individual cell/mL cell, cell quantity expands about 8 times.
Wherein, the total amount of described EDTA-PBS solution should be at (50-100) mL/g Cytodex3.
The method of large-scale culture human umbilical cord mesenchymal stem cells of the present invention adds between umbilical cord in glass culture bottle
Mesenchymal stem cell media, at humidity 95%, 5%CO2Incubator in balance 30 minutes in 35.5 DEG C of incubations；Adjust between umbilical cord
The temperature of mesenchymal stem cell media to 35 DEG C, pH7.05, add microcarrier and the umbilical cord mesenchymal stem cells of pretreatment,
The speed stirring 4h of 10RPM～40RPM；It is warming up to 37 DEG C, in mixing speed, the 5%CO of 35RPM～60RPM2Under the conditions of cultivate
96h, period adds umbilical cord mesenchymal stem cells culture medium.Cultural method of the present invention is simple to operate, safe and effective.Experiment table
Bright, compared with the conventional method, obtain cell after the method propagation of large-scale culture human umbilical cord mesenchymal stem cells of the present invention more,
Cell viability and multiplication capacity are strong, and can keep the form of umbilical cord mesenchymal stem cells and good stem cell properties very well, suitable
Mass propgation for umbilical cord mesenchymal stem cells.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
In having technology to describe, the required accompanying drawing used is briefly described.
Fig. 1 shows the result figure of each surface marker of cell after embodiment 5 Flow cytometry cryopreservation resuscitation, and wherein figure a is
Cell sign CD14, figure b are CD44, figure c is CD45, figure d is CD29, figure e is CD90, figure f is CD105.
Detailed description of the invention
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only a part of embodiment of the present invention rather than whole embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under not making creative work premise, all
Belong to the scope of protection of the invention.
In order to further illustrate the present invention, below in conjunction with specific embodiment, the present invention will be described in detail.Wherein, each reality
Execute the operation in example all to carry out under rigorous aseptic environment.
Embodiment 1, umbilical cord mesenchymal stem cells separation and Extraction
Take people's umbilical cord and isolate China's Tong Shi glue, shred to (1-5) mm3After size, at the cell culture fluid containing serum FBS
In culture vessel, in 37 DEG C, 5%CO2Under the conditions of quiescent culture, fluid infusion first after 5～7 days, climb out of in flakes in observing ware
Cell after, sop up culture medium and piece of tissue in ware, carry out changing liquid and process, cultivate 2～3 days, obtain primary umbilical cord mesenchyma dry
Cell.
Wherein, the formula of the described cell culture fluid containing serum FBS is DMEM/F12+15%FBS；The formula of described fluid infusion
For DMEM/F12+15%FBS.
Embodiment 2, the culture vessel of umbilical cord mesenchymal stem cells cultivate amplification
When primary umbilical cord mesenchymal stem cells covers with the 80% of plate, discard original fluid, wash 2 times with PBS, add
Covering the trypsin containing EDTA of amount at the bottom of ware, digest 1-2min, period uninterruptedly observes with inverted microscope, sees that kytoplasm returns
Contracting, intercellular substance increases, cell rounding, and the cell culture medium adding respective volume immediately terminates digestion, repeatedly blows with pipet
Beating attached cell, blown and beaten by cell, 1500r/min is centrifuged 5min, then uses cell culture fluid re-suspended cell, according to density is
6×104Individual cell/mL, is inoculated in plate, adds fresh cell medium in 37 DEG C, 5%CO2Under the conditions of cultivate, within 3 days, change
Liquid, obtains P1 for cell.I.e. can get 2 × 105Individual cell/mL cell, cell quantity expands about 3 times.
Embodiment 3, umbilical cord mesenchymal stem cells cultivate amplification in containing microcarrier bioreactor
1. glass culture bottle pretreatment: working volume is that the glass culture bottle of 500mL cleans up, dries to completely dry
Dry, addition silication liquid is in bottle, and slow rotating and culturing bottle makes silication immersion moisten bottle wall, after sucking silication liquid, is placed in by rolling bottle logical
Air-drying 12h at wind, distilled water flushing is standby.
2. microcarrier pretreatment: dry microcarrier Cytodex-3 is without Ca2+、Mg2+, pH 7.4 phosphate buffer in
At room temperature soak and expand at least 3 hours, abandoning supernatant, with new preparation without Ca2+、Mg2+, pH 7.4 PBS wash micro-load
Body several minutes, discards PBS, changes new without Ca2+、Mg2+, the PBS of pH 7.4, then microcarrier solution autoclaving is gone out
Bacterium；Before use, first allowing aseptic microcarrier settle, incline supernatant, then with the culture medium rinse of warm, then allows microcarrier
Sedimentation, supernatant discarded, then resuspended rear standby by a small amount of culture medium.
3. the inoculation of umbilical cord mesenchymal stem cells: add the cell culture medium of 1/3-1/2 final volume in glass culture bottle,
Humidity 95%, 5%CO2Incubator in balance 30 minutes in 35.5 DEG C of incubations；Adjust the temperature to 35 DEG C of cell culture medium,
PH7.05, after the microcarrier of addition pretreatment is 0.5mg/mL-5.0mg/mL to concentration, is slowly added to primary umbilical cord mesenchyma and does
Cell to concentration is 0.5 × 105cells/mL-2×105Cells/mL, stirs under lucifuge with the mixing speed of 10RPM～40RPM
Mixing 4 hours, within an hour, homogeneous temperature rises to 37 DEG C afterwards, mixing speed is promoted to 35～60RPM, adds cell training
Foster base is to final volume, in 37 DEG C, 5%CO2Under the conditions of cultivate.Every 48h changes the culture medium of 50%,.
4. the acquisition of umbilical cord mesenchymal stem cells: stop stirring, microcarrier settles, and supernatant discarded culture medium, with pH7.6's
Containing EDTA without Ca2+、Mg2+The PBS washing of ion, the total amount of EDTA-PBS solution should be at (50-100) mL/g
Cytodex3.Then use pancreas enzyme-EDTA solution digestion, after 15min, add the culture fluid containing 10% (v/v) serum substitute
Terminate digestion, obtain P2 for cell, available 1.6 × 106Individual cell/mL cell, cell quantity expands about 8 times.
Embodiment 4, the freezen protective of umbilical cord mesenchymal stem cells
Plate will cover with the umbilical cord mesenchymal stem cells to 80-90%, remove old culture fluid, with PBS 2 times,
It is subsequently added into the pancreatin 37 DEG C digestion 1-2min containing EDTA, after cell all comes off, adds cell culture fluid and terminate digestion,
1500rpm/min is centrifuged 5min, removes supernatant, then with stem cell cryopreserving liquid, cell is resuspended, it is thus achieved that umbilical cord mesenchyma dry thin
Born of the same parents' suspension joins in cell cryopreservation tube, be placed in the program mode cooling box containing isopropanol-80 DEG C overnight, within second day, transfer to-
The medium-term and long-term preservation of liquid nitrogen of 196 DEG C.
Embodiment 5, flow cytometry inspection recovery cell surface marker
Take 1 pipe freeze-stored cell 37 DEG C recovery after add umbilical cord mesenchymal stem cells culture medium (DMEM/F12+15%FBS) in
37 DEG C, 5%CO2Under the conditions of cultivate and gather in the crops 3 × 10 after 3 days6Cell, as flow cytometer detection.Testing result is as shown in Figure 1.
Result shows, high expressed mescenchymal stem cell mark CD29, CD44 and CD105 that this cell is homogeneous, expresses dry
Cell sign CD90, does not express hematopoietic stem cell mark CD14 and CD45, and this identifies knot with mescenchymal stem cell surface antigen
The most consistent, show that the umbilical cord mesenchymal stem cells after cryopreservation resuscitation still keeps the surface character of mescenchymal stem cell.
Claims (8)
1. a method for large-scale culture human umbilical cord mesenchymal stem cells, comprises the steps:
1) in glass culture bottle, umbilical cord mesenchymal stem cells culture medium is added, at humidity 95%, 5%CO2Incubator in
35.5 DEG C of incubations balance 30 minutes；
2) adjust the temperature to 35 DEG C of umbilical cord mesenchymal stem cells culture medium, pH7.05, add microcarrier and the umbilical cord of pretreatment
Mescenchymal stem cell, the speed stirring 4h of 10RPM～40RPM；
3) 37 DEG C it are warming up to, in mixing speed, the 5%CO of 35RPM～60RPM2Under the conditions of cultivate 96h, period adds and fills between umbilical cord
Matter stem cell media.
Method the most according to claim 1, the addition of the microcarrier of described pretreatment is final concentration of to microcarrier
0.5mg/mL-5.0mg/mL。
Method the most according to claim 1 and 2, described microcarrier be Cytodex1 microcarrier, Cytodex2 microcarrier,
Cytodex3 microcarrier, Cytopore microcarrier or Cytoline microcarrier.
4., according to the method described in claim 1-3 any one, the method for described microcarrier pretreatment is that microcarrier first uses pH
The phosphate buffer of 7.4 soaks and expands, sterilizing；Before using, with a small amount of after the culture medium rinse of aseptic microcarrier warm
Culture medium is resuspended.
5., according to the method described in claim 1-4 any one, the addition of described umbilical cord mesenchymal stem cells is to umbilical cord
Mescenchymal stem cell final concentration of 0.5 × 105cells/mL-2×105cells/mL。
6. according to the method described in claim 1-5 any one, step 2) described stirring carries out under lucifuge.
7. according to the method described in claim 1-6 any one, described in add umbilical cord mesenchymal stem cells culture medium be specially
Every 48h changes the culture medium of 50%.
8., according to the method described in claim 1-7 any one, the formula of described umbilical cord mesenchymal stem cells culture medium is
DMEM/F12+15%FBS.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107653226A (en) * | 2017-11-15 | 2018-02-02 | 南京三生生物技术股份有限公司 | A kind of human umbilical cord mesenchymal stem cells isolated culture method |
| CN109706213A (en) * | 2018-12-28 | 2019-05-03 | 广州赛莱拉干细胞科技股份有限公司 | A kind of method of quick screening cell microcarrier culture systems |
| CN110093312A (en) * | 2019-05-15 | 2019-08-06 | 张永国 | A kind of cell large-scaled culture method, purification process and cell messenger |
| CN110804587A (en) * | 2019-11-27 | 2020-02-18 | 浙江卫未生物医药科技有限公司 | Method for modeling production of mesenchymal stem cells by adopting improved microcarrier cell culture method |
| CN111139220A (en) * | 2020-01-08 | 2020-05-12 | 深圳市旷逸生物科技有限公司 | Three-dimensional culture method of umbilical cord mesenchymal stem cells |
| WO2020173280A1 (en) * | 2019-02-28 | 2020-09-03 | 京东方科技集团股份有限公司 | Preparation method for umbilical cord mesenchymal stem cells and cell sheets thereof |
| CN113621566A (en) * | 2021-06-16 | 2021-11-09 | 广州赛莱拉干细胞科技股份有限公司 | Umbilical cord mesenchymal stem cell separation culture method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220338A (en) * | 2011-05-06 | 2011-10-19 | 武汉北度生物科技有限公司 | Method for expressing human 2.5 S beta-nerve growth factor in human umbilical cord mesenchymal stem cell, and separating and purifying the human 2.5 S beta-nerve growth factor |
| CN102321566A (en) * | 2011-09-13 | 2012-01-18 | 协和干细胞基因工程有限公司 | Method for extensive amplification of cell line and mesenchymal stem cell in vitro |
| WO2012142569A2 (en) * | 2011-04-15 | 2012-10-18 | The Regents Of The University Of California | Decellularized extracellular matrix |
-
2016
- 2016-11-24 CN CN201611051086.6A patent/CN106318906A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012142569A2 (en) * | 2011-04-15 | 2012-10-18 | The Regents Of The University Of California | Decellularized extracellular matrix |
| CN102220338A (en) * | 2011-05-06 | 2011-10-19 | 武汉北度生物科技有限公司 | Method for expressing human 2.5 S beta-nerve growth factor in human umbilical cord mesenchymal stem cell, and separating and purifying the human 2.5 S beta-nerve growth factor |
| CN102321566A (en) * | 2011-09-13 | 2012-01-18 | 协和干细胞基因工程有限公司 | Method for extensive amplification of cell line and mesenchymal stem cell in vitro |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107653226A (en) * | 2017-11-15 | 2018-02-02 | 南京三生生物技术股份有限公司 | A kind of human umbilical cord mesenchymal stem cells isolated culture method |
| CN109706213A (en) * | 2018-12-28 | 2019-05-03 | 广州赛莱拉干细胞科技股份有限公司 | A kind of method of quick screening cell microcarrier culture systems |
| WO2020173280A1 (en) * | 2019-02-28 | 2020-09-03 | 京东方科技集团股份有限公司 | Preparation method for umbilical cord mesenchymal stem cells and cell sheets thereof |
| CN110093312A (en) * | 2019-05-15 | 2019-08-06 | 张永国 | A kind of cell large-scaled culture method, purification process and cell messenger |
| CN110804587A (en) * | 2019-11-27 | 2020-02-18 | 浙江卫未生物医药科技有限公司 | Method for modeling production of mesenchymal stem cells by adopting improved microcarrier cell culture method |
| CN111139220A (en) * | 2020-01-08 | 2020-05-12 | 深圳市旷逸生物科技有限公司 | Three-dimensional culture method of umbilical cord mesenchymal stem cells |
| CN113621566A (en) * | 2021-06-16 | 2021-11-09 | 广州赛莱拉干细胞科技股份有限公司 | Umbilical cord mesenchymal stem cell separation culture method |
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