Culture method for inducing differentiation of umbilical cord mesenchymal stem cell into cartilage and culture medium used in method
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A mesenchymal stem cell and culture medium technology, applied in the field of cell induction culture, can solve the problems of low differentiation rate, fast dedifferentiation speed, inability of mesenchymal stem cells to differentiate into chondrocytes, etc., and achieve the effect of increasing differentiation rate and induction speed
Inactive Publication Date: 2019-03-12
BEIJING TAIDONG BIOTECH CO LTD
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Problems solved by technology
The existing induction technology has a relatively low induction differentiation rate and a long time period. It takes about 14 to 28 days to form cartilage, and it cannot effectively differentiate all mesenchymal stem cells into chondrocytes, resulting in low density of induced cartilage, which is not enough. Dense, poor cartilage quality, fast dedifferentiation, unfavorable for future use in tissue engineering
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Embodiment 1
[0045] 1. Inducing components
[0046] name
components
TGF-β3
7ng / mL
Dexamethasone
0.07μmol / L
Ascorbate
40μg / mL
1×ITS
1.3％
L-proline
30μg / mL
sodium pyruvate
1.3mmol / L
[0047] 2. Preparation of induction medium
[0048] 1) The basal medium for inducing differentiation is DMEM high-glucose medium.
[0049]2) Calculate the amount of induction components TGF-β3, dexamethasone, ascorbate, 1×ITS, L-proline, and sodium pyruvate by preparing 100 mL of complete induction medium, and pack them in sterile 0.5 mL EP tubes. Store at -20°C. All components were protected from light.
[0050] 3) When preparing, take out the required volume of DMEM high-glucose medium, dexamethasone, ascorbate, 1×ITS, L-proline, sodium pyruvate, dissolve and mix to make a premix, which can be stored at 4°C 5-7d.
[0051] 4) Before use, add TGF-β3 to make a complete medium, and try to use up the complete medium within 24 ho...
Embodiment 2
[0066] 1. Inducing components
[0067] name
components
TGF-β3
13ng / mL
Dexamethasone
0.13μmol / L
Ascorbate
60μg / mL
1×ITS
0.7％
L-proline
50μg / mL
sodium pyruvate
0.7mmol / L
[0068] 2. Preparation of induction medium
[0069] With embodiment 1.
[0070] 3. Induction of Differentiation Steps
[0071] With embodiment 1.
Embodiment 3
[0073] 1. Inducing components
[0074]
[0075]
[0076] 2. Preparation of induction medium
[0077] With embodiment 1.
[0078] 3. Induction of Differentiation Steps
[0079] With embodiment 1.
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Abstract
The invention relates to the technical field of cell induction culture, and particularly relates to a culture method for inducing the differentiation of an umbilical cord mesenchymal stem cell into acartilage, and a culture medium used in the method. The culture medium is obtained by at least additionally adding the following components in a basal culture medium: 7 ng / mL to 13 ng / mL of TGF (Transforming Growth Factor)-beta 3, 0.07 mumol / L to 0.13 mumol / L of dexamethasone, 40 mug / mL to 60 mug / mL of ascorbate, 0.7 w / v% to 1.3 w / v% of 1*ITS (Internal Transcribed Spacer), 30 mug / mL to 50 mug / mL of L-proline and 0.7 mmol / L to 1.3 mmol / L of sodium pyruvate. According to the culture medium, through optimizing an induction component and matching a specific culture method, the induction velocity is improved; the hard and compact cartilage can be formed after 7 days; all inoculated mesenchymal stem cells can be differentiated into cartilage cells, and the differentiation rate is improved. An induced cartilage globule is compact in texture, and displayed through specific staining, and all mesenchymal stem cells are sufficiently differentiated into the cartilage cells.
Description
technical field [0001] The invention relates to the technical field of cell induction culture, in particular to a culture method for inducing chondrogenic differentiation of umbilical cord mesenchymal stem cells and a culture medium used in the method. Background technique [0002] Mesenchymal stem cells (MSCs) are pluripotent stem cells derived from mesoderm with high self-renewal ability and multilineage differentiation potential. In fact, MSCs widely exist in various tissues throughout the body, and almost come from all tissues and organs of the body, such as bone marrow, periosteum, adipose tissue, dental pulp, synovium, umbilical cord, placenta, amniotic fluid and fetal tissue. Mesenchymal stem cells from different sources have similar morphology, express the same surface markers, and have similar biological characteristics. For example, they can be cultured and expanded in vitro, and can differentiate into neurons, osteoblasts, Cells of multiple tissue systems includi...
Claims
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