CN103805562B - Cultivate the serum free medium of placenta mesenchyma stem cell - Google Patents
Cultivate the serum free medium of placenta mesenchyma stem cell Download PDFInfo
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- CN103805562B CN103805562B CN201410018678.2A CN201410018678A CN103805562B CN 103805562 B CN103805562 B CN 103805562B CN 201410018678 A CN201410018678 A CN 201410018678A CN 103805562 B CN103805562 B CN 103805562B
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Abstract
A kind of serum free medium cultivating placenta mesenchyma stem cell, based on DMEM nutrient solution, also containing fibroblast growth factor acceptor 2, tethelin, Regular Insulin, Transferrins,iron complexes, gsh, BMP-4, L-glutaminate, Sodium.alpha.-ketopropionate, non-essential amino acid and beta-mercaptoethanol etc.Various serum free medium provided by the invention can make placenta mesenchyma stem cell growing multiplication in serum free medium system effectively, serum cell is comparatively had to have higher growing multiplication speed, and keep its stem cell properties, there is the potential of multiple differentiation, can directional induction lipoblast and scleroblast etc.
Description
Technical field
The present invention relates to a kind of composition for cell cultures, particularly relate to a kind of composition of culturing stem cells, cultivate placenta mesenchyma stem cell in serum-free mode.
Background technology
Mescenchymal stem cell (Mesenchymal Stem Cells, MSCs) is a type of stem cell, possesses two key characters of stem cell: very strong self duplication ability and many differentiation potentials.MSCs originates from mesoderm, and it can break up to mesoderm tissues theoretically.The mescenchymal stem cell of the mankind is separated the earliest from marrow, in vivo with under external specified conditions there is the ability (J.Orthop.Res. broken up to the multiple adult cell such as scleroblast, chondrocyte, adipocyte, endotheliocyte, neurocyte, myocyte, liver cell, 1991,641-650; Science, 1999,28,143-147).
Up-to-date research shows that mescenchymal stem cell has immunomodulatory and Hematopoiesis Support affect, and is easy to foreign gene importing expression.Therefore mescenchymal stem cell not still tissue-engineered bone, cartilage and cardiac muscle build in seed cell, carrier cell important in gene therapy, and due to mescenchymal stem cell promotion hematopoietic reconstitution and the anti-host response function of inhibition of transplant, be with a wide range of applications in hematopoietic stem cell transplantation and organ transplantation.Mescenchymal stem cell has the characteristic of growth-arrested in vitro, utilizes this characteristic, and people success separation and Culture from the Various Tissues such as liver, kidney, pancreas, muscle, cartilage, skin, peripheral blood go out mescenchymal stem cell.
The mescenchymal stem cell reported at present is mainly derived from marrow, adopts density gradient centrifugation to obtain.Although separation method is easy, donor gets the operation that marrow needs experience one more painful, and has very high infection chance in the process of drawing materials and after drawing materials; Because the content of MSC in human bone marrow is extremely rare, every 10
5~ 10
6approximately only have 1 in individual mononuclearcell, and along with the increase at age, in marrow, the quantity of mescenchymal stem cell, propagation and differentiation capability all significantly decline, and make it be restricted in investigation and application especially clinical application.The placenta originating from embryonic development period extraembryonic mesoderm is made up of interstitial, blood vessel and nurse cell, containing a large amount of mesenchyme compositions.Up-to-date research shows containing abundant stem cell in placenta, and from placenta, separation and Culture goes out these multipotential stem cells and will open up a brand-new and abundant source for experimental study and clinical application.
Chinese invention patent application 200810061267.6 discloses a kind of placenta mesenchyma stem cell and is separated and amplification in vitro cultural method, by collection after the histocyte of placenta parent side decidua, first carry out the adherent amplification of primary cell, the positive and negative method combined to immune sorting go again with purifying placenta mesenchyma stem cell, obtains CD34
-cD105
+cell, subculture amplification cultivation in serum-free culture system in vitro.
Chinese invention patent ZL200910117522.9 discloses a kind of method of separating and extracting stem cells from placenta, umbilical cord or fatty tissue, first by placenta, umbilical cord or fatty tissue and phosphoric acid buffer by 2.5 ~ 4: 1 weight ratio mix to put into and organize grinding barrel, collagenase mixing is added after pulverizing, after hatching under about 37 DEG C conditions, filtering and add precipitation agent, Aspirate supernatant, centrifugal, removing supernatant liquor, joins Sodium Diatrizoate-ficoll 400 by concentrated cell
#liquid, more centrifugal, collect the cellular layer of middle 10-15ml, with the cleaning of cell maintenance liquid, can use after qualified for the cell counting of collection.
Chinese invention patent application 201110005964.1 discloses a kind of method extracting primary mesenchymal stem cells and serum-free amplification from placenta, the fresh miscarriage placenta obtained is carried out bloodletting immediately, adopts physiological buffer to remove hyaluronic mucus and female blood stains dye of placenta tissue and umbilical cord appearance; Use physiological buffer perfusion, to remove hemocyte and fragment of tissue; Shredded by placental lobules, with collagenase digesting, collecting cell composition, by adherent sieve method purifying mesenchymal stem cells.
These technical schemes existing mostly separate stem cells from placenta, thus set up placenta stem-cell storehouse, still there is shortcomings, be mainly reflected in the several respects such as the cell quantity of purity and/or acquisition, so that be still difficult to the expectation meeting people.
In addition, to extract from tissue and after obtaining mescenchymal stem cell, to the cultivation of mesenchymal cell, still need to add foetal calf serum and carry out adherent culture, also be not still applicable to the substratum of industry application, this adds cost and risk in cost and follow-up clinical test, needs to explore further.
Summary of the invention
One object of the present invention is to provide a kind of serum free medium cultivating placenta mesenchyma stem cell, realizes placenta mesenchyma stem cell vitro culture in serum-free mode.
Another object of the present invention is to provide a kind of method of cultivating placenta mesenchyma stem cell with serum free medium, effectively to improve the in-vitro separation of placenta mesenchyma stem cell and to produce efficiency.
A kind of serum free medium cultivating placenta mesenchyma stem cell provided by the invention, by DMEM(Dulbeco ' s Modified Eagle Medium) based on nutrient solution, also containing fibroblast growth factor acceptor 2 (Fibroblast Growth Factor2, FGF-2), tethelin, Regular Insulin, Transferrins,iron complexes (TF), gsh, bone morphogenetic protein 4 (Morphogenetic protein-4, and L-glutaminate, Sodium.alpha.-ketopropionate (sodium pyruvate), non-essential amino acid and beta-mercaptoethanol etc. BMP-4).
Another kind provided by the invention cultivates the serum free medium of placenta mesenchyma stem cell, by DMEM(Dulbeco ' s Modified Eagle Medium) based on nutrient solution, also containing concentration 10 μ g/ml Regular Insulin, 0.1 μ g/ml Transferrins,iron complexes, 0.1ng/mlBMP-4,10ng/mlFGF-2 and 3ng/ml tethelin.
Another kind provided by the invention cultivates the serum free medium of placenta mesenchyma stem cell, by DMEM(Dulbeco ' s Modified Eagle Medium) based on nutrient solution, also containing concentration 10 μ g/ml Regular Insulin, 0.1 μ g/ml Transferrins,iron complexes, 200 μ g/ml gsh, 0.1ng/mlBMP-4,10ng/mlFGF-2 and 3ng/ml tethelin.
Another kind provided by the invention cultivates the serum free medium of placenta mesenchyma stem cell, by DMEM(Dulbeco ' s Modified Eagle Medium) based on nutrient solution, also containing concentration 10 μ g/ml Regular Insulin, 0.1 μ g/ml Transferrins,iron complexes, 200 μ g/ml gsh, 0.1ng/mlBMP-4,10ng/mlFGF-2 and 3ng/ml tethelin, 2mML-glutamine, concentration 55 μMs of beta-mercaptoethanols, concentration 1mM Sodium.alpha.-ketopropionate and concentration 1v/v% non-essential amino acid.
In the present invention, non-essential amino acid is L-glutamic acid, L-Ala, glycine, aspartic acid, Gelucystine, proline(Pro), Serine and tyrosine etc., obtains by business-like approach, as: but be not limited only to Gibco company etc.It will be appreciated by those skilled in the art that the non-essential amino acid used as substratum is interpreted as the mixture of above-mentioned 8 seed amino acids.
The mescenchymal stem cell that the above-mentioned various serum free medium of the present invention is applicable to taking from placenta carries out succeeding transfer culture, realizes the adherent growth of mescenchymal stem cell, and keeps the multiplication capacity of cell.
A kind of method of cultivating placenta mesenchyma stem cell with serum free medium provided by the invention, get the chorion of the denuded amniotic membrane layer of placenta fetus side, shred the digestion of rear 0.13v/v% II Collagenase Type and collecting cell, adherent culture is carried out with DMEM substratum (containing 10% foetal calf serum and 2mML-glutamine), obtain the mescenchymal stem cell of adherent growth, then carry out body succeeding transfer culture with the above-mentioned various serum free medium of the present invention.
Empirical tests, various serum free medium provided by the invention can make placenta mesenchyma stem cell growing multiplication in serum free medium system effectively, serum cell is comparatively had to have higher growing multiplication speed, and keep its stem cell properties, there is the potential of multiple differentiation, can directional induction lipoblast and scleroblast etc.
The beneficial effect that technical solution of the present invention realizes:
The serum free medium of cultivation placenta mesenchyma stem cell provided by the invention, by studying various somatomedin kind and various ingredients and consumption and obtain, achieves placenta mesenchyma stem cell vitro culture in serum-free mode.In this serum free medium system, mescenchymal stem cell can realize adherent growth and effectively improve the amplification ability of mescenchymal stem cell, makes the culture system not containing external source serum more be conducive to stem cell and is applied to clinical transplantation safely.
The present invention also effectively improves the in-vitro separation of placenta mesenchyma stem cell and produces efficiency, and serum free medium is directly applied to obtained placenta mesenchyma stem cell, and the mescenchymal stem cell quantity of acquisition is significantly improved.
Accompanying drawing explanation
Fig. 1 is the growing state of separated primary mescenchymal stem cell in the substratum containing serum; Wherein, Figure 1A is that cell is fusiformis under inverted microscope (10 × 10 times); Figure 1B is that cell inoculation forms cell monolayer in latter 3 days-5 days;
Fig. 2 is that separated primary mesenchymal cell is first after primary adherent culture in the substratum containing serum, then the growing state figure in serum free medium, uses inverted microscope, 10 × 10 times of observations;
Fig. 3 is for have blood serum medium and serum free medium to carry out amplification ability comparative analysis result figure to placenta mesenchyma stem cell to employing;
Fig. 4 A adopts the mescenchymal stem cell that goes down to posterity of serum free medium of the present invention, through differentiation-inducing and be stearoblast, its oil red O stain result figure; Wherein, arrow " 1 " represents that red fat drips; Arrow " 2 " represents blue cell core;
Fig. 4 B adopts the mescenchymal stem cell that goes down to posterity of serum free medium of the present invention, through differentiation-inducing and be scleroblast, its alkaline phosphatase staining result figure; Wherein, arrow " 3 " represents dark-brown or aterrimus endochylema.
Embodiment
Technical scheme of the present invention is described in detail below in conjunction with accompanying drawing.The embodiment of the present invention is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to the technical scheme of invention or equivalent replacement, and not departing from the spirit and scope of technical solution of the present invention, it all should be encompassed in right of the present invention.
If other reagent unexplained reference used of the embodiment of the present invention, all purchased from Sigma-Aldrich (Sigma-Aldrich) company.
The combination experiment of cytokine in embodiment 1 serum free medium
Selected somatomedin: recombinant human fibroblast growth factor 2(hFGF), biosynthetic human insulin (I), recombinant human growth hormone (hGH), people's recombinant transferrin (TF) and BMP-4 concentrated solution be mixed with concentrated solution, through aseptic 0.22 μm of aqueous phase film or organic phase membrane filtration degerming (if this step buys aseptic concentrated solution, not needing to carry out), refrigeration or cold storage for subsequent use.
Different concentration (see table 1) is designed for different somatomedins to add in nutrient solution and the optimum effect concentration finding this somatomedin according to growth curve.The different somatomedins of optimum concn carry out combining (see table 2) by the screening experiment through over-richness, investigate the impact (see table 3) separately on stem cell growth situation, obtain optimum growh combinations of factors nutrient solution.
The concentration of each somatomedin of table 1
The different combination group of somatomedin of table 2 combination experiment and the concentration of somatomedin
D1 and D2 all represents null term, the numerical markings that " 1 " and "-1 " is orthogonal experiment.
The contrast of growth and proliferation of cell in the substratum system of table 3 various combination
Interpretation: each somatomedin optimum effect concentration is finally determined, the optimum concn of the optimum concn of hGH to be the optimum concn of 3ng/ml, hFGF be 10ng/ml, TF is 0.1 μ g/ml, the optimum effect concentration of Regular Insulin is the optimum effect concentration of 10 μ g/ml, BMP4 is 0.1ng/ml.
Embodiment 2
Get the chorion of placenta fetus side, shred, collecting cell after the digestion of 0.13v/v% II Collagenase Type, carries out adherent culture with cell culture medium (containing DMEM, 10v/v% foetal calf serum, 2mML-glutamine).
1. cell carries out adherent culture in culture dish, discards substratum and suspension cell after 48h.Every 2-3 days changes liquid once, treats that cell confluency reaches 90%, with Tryspin-EDTA digestion and harvested cell, the cells rinsed with PBS of results once after, then renewed vaccination, and add fresh culture and carry out Secondary Culture.
2. the cell of results is carried out succeeding transfer culture, used medium is liquid based on DMEM, also containing 2mML-glutamine, 55uMb-mercaptoethanol, 1mM Sodium.alpha.-ketopropionate, 1v/v% non-essential amino acid (Gibco company), 10 μ g/ml Regular Insulin, 0.1 μ g/ml Transferrins,iron complexes, 200 μ g/ml gsh, 0.1ng/mlBMP-4,10ng/mlFGF-2 and 3ng/ml hGH) serum free medium.
Embodiment 3
For the mensuration of surface markers having the cell cultivating acquisition in serum and serum free medium, the different culture system of Flow Cytometry Assay is adopted to cultivate the ratio (see table 3) of different markup cell-surface antigens in the mescenchymal stem cell of results.
Table 3
CD90 in primary placenta fetus side chorionic cells
+, CD73
+and CD105
+cell is average out to 99.06%, 99.98% and 99.10% respectively, shows that the cambiform cell of primary adherent has significant mesenchymal cell characteristic.Cultivate through 2 passaging anchorage, having in blood serum medium, CD90
+and CD105
+cell has remarkable minimizing, and in serum free medium CD90
+and CD105
+cell still can exist with higher ratio, maintains the characteristic of mescenchymal stem cell.According to above-mentioned comparison, can show that from the chorion of placenta fetus side, be separated the cell process in serum free medium obtained goes down to posterity, and has mescenchymal stem cell surface antigen feature.
Embodiment 4
Observe the cellular form that different culture system obtains.See Fig. 2 and Fig. 3, the placenta fetus side chorion histocyte of II Collagenase Type digestion can see spindle shape cell attachment in 24 hours after inoculation, and in radiating or concentric circles arrangement.A large amount of colonies appeared at inoculation after 5 days, then laying bottom culture dish gradually.In 100mm culture dish, cell amplification forms individual layer needs 7 days-10 days time.Under Fig. 1 A is presented at inverted microscope, cell is fusiformis, 10 × 10 times.After cell goes down to posterity through 2 times, cell is significant fibrous cell, forms cell monolayer (see Fig. 1 B) after cell inoculation through 3 days-5 days.After primary adherent is cultivated, cell carries out amplification Secondary Culture in serum free medium, obtains in best serum free culture system through overtesting, and in 100mm culture dish, cell forms cell monolayer in 2 days-3 days, and cell is fiber shape (see Fig. 3).
Embodiment 5 increases ability comparative analysis
Blood serum medium and serum free medium is had to carry out the comparative analysis of amplification ability to placenta mesenchyma stem cell to employing.Gather in the crops through the primary and cell of 2 times of going down to posterity, the cell of two kinds of different culture systems inoculates 24 orifice plates, and the amplification ability of carrying out cell compares, and the Dynamic Graph that cell quantity increases is see Fig. 4.
From the comparative result analysis of Fig. 4, in the 1st day-8 days that cultivate, the cell under two kinds of culture systems has growing multiplication, and the cultivation effect of serum free medium is more obvious.
Embodiment 6 stem cell confirms
Placenta mesenchyma stem cell is to adipocyte and osteoblastic differentiation-inducing confirmation.
Be taken in serum free culture system the placenta mesenchyma stem cell cultivating for 2 generations, be prepared into single cell suspension, with 1 × 10
4individual/hole is inoculated in 6 orifice plates carries out adherent culture, and 24h adipocyte inducing culture (purchased from Gibco) carries out the Induction of committed differentiation of adipocyte; With 5 × 10
3individual/hole is inoculated in 6 orifice plates carries out adherent culture, and 24h osteoblast induction medium (purchased from Gibco) carries out osteoblastic Induction of committed differentiation.
In the experiment of adipocyte Induction of committed differentiation, under inverted microscope, have circular fat to drip calmness in visible cell, cell appearance becomes circle, and oval, nucleus is dripped by fat and squeezes in side.Oil red O stain showed cell core is in blue, and fat drips take on a red color (see Fig. 4 A), and along with the prolongation of induction time, the ratio of adipocyte increases gradually.
In the experiment of scleroblast Induction of committed differentiation, can see that cellular form changes cube into, refractivity is strong, and along with the prolongation of induction time, cuboidal cell proportion increases gradually.Cell continues propagation, forms cladding.Be dark-brown or aterrimus (participating in Fig. 4 B) with the inducing cell endochylema of alkaline phosphatase staining, be wherein full of fine and close black precipitate.
Claims (3)
1. cultivate the serum free medium of placenta mesenchyma stem cell for one kind, it is characterized in that based on DMEM nutrient solution, also containing concentration 10 μ g/ml Regular Insulin, 0.1 μ g/ml Transferrins,iron complexes, 0.1ng/ml BMP-4,10ng/ml FGF-2,3ng/ml tethelin, 200 μ g/ml gsh, 2mM L-glutaminate, concentration 55 μMs of beta-mercaptoethanols, concentration 1mM Sodium.alpha.-ketopropionate and concentration 1v/v% non-essential amino acid.
2. the serum free medium of cultivation placenta mesenchyma stem cell according to claim 1, is characterized in that described non-essential amino acid is L-glutamic acid, L-Ala, glycine, aspartic acid, Gelucystine, proline(Pro), Serine and tyrosine composition.
3. cultivate the method for placenta mesenchyma stem cell with serum free medium for one kind, get the chorion of the denuded amniotic membrane layer of placenta fetus side, shred the digestion of rear 0.13v/v% II Collagenase Type and collecting cell, to carry out adherent culture containing the DMEM substratum of 10% foetal calf serum and 2mM L-glutaminate, obtain the mescenchymal stem cell of adherent growth, then carry out body succeeding transfer culture with the serum free medium described in claim 1 or 2.
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Families Citing this family (29)
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| CN112662623A (en) * | 2021-01-25 | 2021-04-16 | 中科博生生物工程有限公司 | Serum-free stem cell culture medium and application thereof |
| CN114958734B (en) * | 2021-10-22 | 2023-03-24 | 中国医学科学院生物医学工程研究所 | Human mesenchymal stem cell culture medium |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2272839C2 (en) * | 1998-11-09 | 2006-03-27 | Консорцио Пер Ла Джестионе Дель Чентро Ди Биотекнолоджие Аванцате | Serum-free medium for cultured cells used for reconstruction of osseous and cartilage segments (variants) |
| CN101270349A (en) * | 2008-03-20 | 2008-09-24 | 浙江大学 | Method for placenta mesenchyma stem cell separation and in vitro amplify cultivation |
| KR20130112028A (en) * | 2010-06-17 | 2013-10-11 | 스템알디, 인크. | Serum-free chemically defined cell culture medium |
| CN102146359A (en) * | 2011-01-13 | 2011-08-10 | 王泰华 | Method for extracting original mesenchymal stem cells from placenta and serum-free amplification |
| CN102732477B (en) * | 2012-06-15 | 2013-06-19 | 江苏瑞思坦生物科技有限公司 | Human adipose-derived stem cell serum-free basic medium |
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