Cell transplantation culture method for forming bone tissue in vivo

A technology of cell transplantation and culture method, which is applied in the field of osteoblast-like cells and cell transplantation and culture, and can solve the problems of poor plasticity, poor degradation performance and slow degradation of osteogenic tissue blocks.

Inactive Publication Date: 2015-12-23
YUNNAN JICI INSITUTE FOR REGENERATIVE MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many biomaterials currently in use have a fixed structure and slow degradation in the body. After transplanting cells, the structural plasticity of the osteogenic tissue block is poor, and the properties such as extensibility and compression resistance are far inferior to their own bone tissue; at the same time , the degradation performance is poor, and it will also affect other tissues

Method used

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  • Cell transplantation culture method for forming bone tissue in vivo
  • Cell transplantation culture method for forming bone tissue in vivo
  • Cell transplantation culture method for forming bone tissue in vivo

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Prepare BDMatrigel to form the required three-dimensional culture matrix

[0022] The stock solution of BDMartigel (concentration 8mg / ml) was taken out from -20°C, equilibrated overnight at 4°C and then used on ice. According to the growth conditions required by the transplanted cells, other required growth factors, proteins or small molecular compounds are added in a certain proportion.

[0023] 2. Preparation of Osteogenic Seed Cells for Transplantation

[0024] In this experiment, the osteogenic seed cells used were derived from human skin fibroblast-derived inducible osteoblasts, and the suitable culture medium for the cells mainly contained compounds such as 9 μM CHIR99021, 10 μM Forskolin and 10 nM dexamethasone. The total number of cells prepared is 2.7x10 6 , the digested cells were centrifuged and dissolved in 50 μl of culture medium containing the above substances;

[0025] 3. Three-dimensional complex of cells and matrix

[0026] Operate on ice at 2-8°...

Embodiment 2

[0033] 1. Prepare BDMatrigel to form the required three-dimensional culture matrix

[0034] The stock solution of BDMartigel (concentration 8mg / ml) was taken out from -20°C, equilibrated overnight at 4°C and then used on ice. According to the growth conditions required by the transplanted cells, other required growth factors, proteins or small molecular compounds are added in a certain proportion.

[0035] 2. Preparation of Osteogenic Seed Cells for Transplantation

[0036] In this experiment, the osteogenic seed cells used were derived from human skin fibroblast-derived inducible osteoblasts, and the suitable culture medium for the cells mainly contained compounds such as 9 μM CHIR99021, 10 μM Forskolin and 10 nM dexamethasone. The total number of cells prepared is 2.7x10 6 , the digested cells were centrifuged and dissolved in 75 μl of culture medium containing the above substances;

[0037] 3. Three-dimensional complex of cells and matrix

[0038] Operate on ice at 2-8°...

Embodiment 3

[0044]1. Prepare BDMatrigel to form the required three-dimensional culture matrix

[0045] The stock solution of BDMartigel (concentration 8mg / ml) was taken out from -20°C, equilibrated overnight at 4°C and then used on ice. According to the growth conditions required by the transplanted cells, other required growth factors, proteins or small molecular compounds are added in a certain proportion.

[0046] 2. Preparation of Osteogenic Seed Cells for Transplantation

[0047] In this experiment, the osteogenic seed cells used were derived from human skin fibroblast-derived inducible osteoblasts, and the suitable culture medium for the cells mainly contained compounds such as 9 μM CHIR99021, 10 μM Forskolin and 10 nM dexamethasone. The total number of cells prepared is 2.7x10 6 , the digested cells were centrifuged and dissolved in 20 μl of culture medium containing the above substances;

[0048] 3. Three-dimensional complex of cells and matrix

[0049] Operate on ice at 2-8°C...

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Abstract

The invention relates to a cell transplantation culture method for forming bone tissue in vivo. The method is characterized in that a three-dimensional matrix BD Matrigel with fast degradation performance is employed to provide a three-dimensional structure for osteogenous seed cells to grow in vivo, so that an osteoid tissue structure can form in osteogenous seed cells. At the same time, BD Matrigel has certain dissolving capacity, corresponding growth factors or small molecular compounds and the like can be added at the early stage of cell transplantation so as to provide a microenvironment suitable for cell differentiation and survival at the early stage of in-vivo growth of the transplanted cells. The method provided by the invention can also be used for transplantation of various cells in tissue engineering, brings convenience to basic research, and brings prospects for clinical application.

Description

technical field [0001] The invention relates to a cell transplantation and culture method for forming bone tissue in vivo, in particular, a bioabsorbable matrix is ​​used to inoculate transplanted osteoblast-like cells, so as to promote the self-organized growth of the cells into bone-like tissue in vivo. It belongs to the field of cell biology and tissue engineering. Background technique [0002] Bone tissue engineering technology co-cultures biomaterials or artificial materials with a specific three-dimensional structure and osteogenic seed cells to form a composite bone tissue material with certain osteogenic plasticity and degradability, which can be used to repair bone tissue defects. On the one hand, there are various sources of osteogenic seed cells, including late-differentiated osteoblasts, osteoblastic cell lines, mixed bone marrow cells, or purified mesenchymal stem cells, as well as non-osteogenic cells such as skin fibroblasts. Differentiated osteoblasts. With...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/22
Inventor 胡敏李燕皎
Owner YUNNAN JICI INSITUTE FOR REGENERATIVE MEDICINE CO LTD
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