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Method for mediated differentiation of bone marrow mesenchyml stem cells into osteoblasts

A technology for osteoblast differentiation and bone marrow mesenchymal, which can be used in bone/connective tissue cells, biochemical equipment and methods, animal cells, etc., and can solve the problem that the efficiency of mediating osteoblast differentiation is not very high.

Inactive Publication Date: 2019-10-22
JIAXING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods require complex material preparation processes, and the efficiency of mediating osteoblast differentiation is still not very high

Method used

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  • Method for mediated differentiation of bone marrow mesenchyml stem cells into osteoblasts
  • Method for mediated differentiation of bone marrow mesenchyml stem cells into osteoblasts
  • Method for mediated differentiation of bone marrow mesenchyml stem cells into osteoblasts

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Bone marrow mesenchymal stem cells (5×10 4 / cm 2 Density), using DMEM medium (10% fetal bovine serum), placed in a cell culture incubator for 24h (conventional culture conditions: 37 ° C, 5% CO 2 concentration) to make the cells fully adhere; then place the culture bottle on a rotating platform to achieve a smooth counterclockwise rotation at 12 to 16 revolutions per minute (preferably 16 revolutions per minute), and then place the whole in the cell culture incubator Culture (conventional culture conditions: 37 ° C, 5% CO 2 concentration), the medium was changed every 2 days; osteoblasts were obtained after 21 days of culture.

[0022] Routine static culture under the same conditions was used as the control group.

[0023] The expression of the osteogenic differentiation index protein alkaline phosphatase (ALP) of osteoblasts was detected, and it was found that the expression of ALP in the osteoblasts cultivated by the method of the present invention was significantl...

Embodiment 2

[0025] Bone marrow mesenchymal stem cells (5×10 4 / cm 2 Density), using DMEM medium (10% volume fraction fetal bovine serum), placed in the cell culture incubator for 24h (conventional culture conditions: 37 ° C, 5% CO 2 concentration) to make the cells fully adhere; then put the culture bottle on a rotating platform to achieve a smooth clockwise rotation at 5 revolutions per minute, and place the whole in a cell culture incubator for culture (conventional culture conditions: 37°C, 5 %CO 2 concentration), the medium was changed every 2 days; after 21 days of culture, the expression of the osteogenic differentiation index protein alkaline phosphatase (ALP) was almost difficult to detect. On the contrary, the expression of adipogenic differentiation index triglyceride (TG) was significantly higher than that of the control group (conventional static culture). see details Figure 4 .

Embodiment 3

[0027] Bone marrow mesenchymal stem cells (5×10 4 / cm 2 Density), using DMEM medium (10% volume fraction fetal bovine serum), placed in the cell culture incubator for 24h (conventional culture conditions: 37 ° C, 5% CO 2 concentration) to make the cells fully adhere; then put the culture bottle on a rotating platform to achieve a smooth anticlockwise rotation at 3 revolutions per minute, and place the whole in a cell culture incubator for culture (conventional culture conditions: 37°C, 5 %CO 2 concentration), the medium was changed every 2 days; after 21 days of culture, the expression of the osteogenic differentiation index protein alkaline phosphatase (ALP) was almost difficult to detect.

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Abstract

The invention discloses a method for mediated differentiation of bone marrow mesenchyml stem cells into osteoblasts. According to the method, based on a conventional cell culturing process, a cell culturing container is simply rotated anticlockwise and is placed in a cell culturing incubator for cell culture, and after 21 days of culture, the osteoblasts can be obtained; especially when the cell culturing container is rotated anticlockwise at the speed of 12-16 revolutions per minute, significant up-regulation of expression of alkaline phosphatase (ALP) of the obtained osteoblasts is achieved.The provided rotation culture method for directional mediated differentiation of the mesenchyml stem cells into the osteoblasts is easy to implement, and under the condition of not adding inducing factors, differentiation of the bone marrow mesenchyml stem cells into the osteoblasts can be significantly improved.

Description

technical field [0001] The invention relates to a method for mediating bone marrow mesenchymal stem cells to differentiate into osteoblasts. Background technique [0002] Stem cell directional differentiation regulation is one of the key topics in stem cell research. Bone marrow mesenchymal stem cells can differentiate into osteoblasts, chondrocytes, fat cells, muscle cells, etc. Mediating the directed differentiation of bone marrow mesenchymal stem cells into osteoblasts to provide suitable cells for transplantation is an urgent problem to be solved in the field of osteoporosis treatment using cell replacement therapy. The conventional method of mediating the directed differentiation of stem cells is by adding various cell-inducing factors, such as dexamethasone and other hormones. These cell-inducing factors are not only expensive, but also easily cause problems such as immunogenicity. Therefore, people try to use other methods to effectively mediate the directed differ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2500/02C12N2500/84
Inventor 唐柏林沈小军董灵庆
Owner JIAXING UNIV
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