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Cell co-culture method for inducing bone mesenchymal stem cells to differentiate into osteoblast

A technology for bone marrow mesenchymal and stem cell differentiation, applied in bone/connective tissue cells, biochemical equipment and methods, tissue culture, etc. Slow and other problems, to achieve the effect of optimizing the co-culture ratio, promoting AP activity, and promoting proliferation

Inactive Publication Date: 2009-06-03
JINAN UNIVERSITY
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Problems solved by technology

[0003] On the issue of inducing bone marrow stromal cells to differentiate into osteoblasts, several existing methods have deficiencies: the conventional induction solution causes the cell growth to slow down, and its specific mode of action and conditions are still controversial; The stimulation of related cell growth factors is still in the exploratory stage, and cell growth factors are used in a large amount, the cost is high, and it can cause adverse reactions in the body, etc.

Method used

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  • Cell co-culture method for inducing bone mesenchymal stem cells to differentiate into osteoblast
  • Cell co-culture method for inducing bone mesenchymal stem cells to differentiate into osteoblast
  • Cell co-culture method for inducing bone mesenchymal stem cells to differentiate into osteoblast

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Embodiment

[0022] 1. Mixed culture of osteoblasts and BMSCs and observation under microscope

[0023] P3 generation OB with 1×10 5 cells / ml were inoculated in 24-well plates, and a control group and 3 experimental groups were set up, a total of 4 groups, with 6 replicate wells in each group. After 24 hours of cell inoculation, the culture medium was discarded, washed twice with PBS, and replaced with serum-free medium. After 24 hours of synchronization, only 1ml of DMEM medium containing 10% fetal bovine serum was added to the control group. Experiments 1, 2, and 3 Add the density to 9×10 respectively 5 / ml, 4×10 5 / ml, 7 / 3×10 5 BMSCs / ml concentration and 1ml DMEM medium containing 10% fetal bovine serum, the ratio of OB and BMSCs to directly co-culture is 1:0, 1:9, 2:8, 3:7. Place the culture plate at 37°C, 5% CO 2 Continue to grow in the incubator. The medium was changed every 3-4 days, and the co-cultivation was continued for 9 days. During the co-culture period, the changes of...

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Abstract

The invention provides a method for inducing bone mesenchymal stem cells to differentiate into osteoblast. The method is characterized in that little osteoblast is added in a bone mesenchymal stem cell culture system to form an osteogenesis micro-environment; the quantity of the osteoblast is more than 30% of the total cell quantity preferably. The method firstly uses the osteogenesis micro-environment provided by little osteoblast, induces the BMSCs to form mature bone tissue in vitro by a co-culture method and has a wide prospect in the field of bone tissue restoration and reconstruction.

Description

technical field [0001] The invention relates to the field of tissue engineering, in particular to a cell co-culture method for inducing bone marrow mesenchymal stem cells (BMSCs) to differentiate into osteoblasts (OB). Background technique [0002] Bone marrow mesenchymal stem cells (BMSCs) can only transform into osteoblasts under specific induction conditions. There are many studies on the conditions for the directed differentiation of BMSCs into osteoblasts. At present, adding chemical substances, cytokines and Methods such as gene transfection are used to achieve the purpose of promoting directed differentiation. [0003] On the issue of inducing bone marrow stromal cells to differentiate into osteoblasts, several existing methods have deficiencies: the conventional induction solution causes the growth of cells to slow down, and its specific mode of action and conditions are still controversial; The stimulation of related cell growth factors is still in the stage of exp...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/08C12N5/077C12N5/0775
Inventor 查振刚林宏生吴昊
Owner JINAN UNIVERSITY
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