Culture method for improving differentiation from peripheral blood multipotent cells to cartilage cells

A technology of pluripotent cells and chondrocytes, applied in the direction of cell culture active agents, biochemical equipment and methods, culture process, etc., can solve the problem of difficult differentiation of stem cells, low efficiency of induction of peripheral blood pluripotent cells, and donor source limited immunity Rejection and disease transmission and other problems, to achieve the effect of promoting proliferation and differentiation, and increasing the rate of differentiation

Inactive Publication Date: 2019-01-01
丰泽康生物医药(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods can repair cartilage tissue damage to a certain extent, but they all have obvious disadvantages. For example, microfracture and subchondral bone drilling methods can make stem cells reach the damaged site, but due to different local environments and mechanical stresses, it is difficult for stem cells to recover. Differentiate into chondrocytes; autologous osteochondral transplantation is safe and effective, but it will bring new trauma to patients; allogeneic osteochondral transplantation avoids ne

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] A culture method for improving the differentiation of peripheral blood pluripotent cells to chondrocytes, comprising the following steps:

[0019] (1) Add sodium citrate anticoagulant to peripheral blood, then add peripheral blood to 0.9% sodium chloride injection, then add 6% hydroxyethyl starch, shake well for 8 minutes, and then let stand 2 hours;

[0020] After layering, absorb the upper layer of cell fluid, and use 1000 rpm to centrifuge for 100 minutes to separate the concentrated solution. The concentrated solution is diluted with normal saline, and then spread on the layered solution prepared by polysucrose and diatrizoate, and then 1000 rpm Centrifuge for 30 minutes, collect the separated stem cell layer, wash the stem cells twice with normal saline, and obtain peripheral blood multipotent mesenchymal stromal cells;

[0021] The peripheral blood multipotential mesenchymal stromal cells are taken for primary culture, and the culture medium used in the primary c...

Embodiment 2

[0029] A culture method for improving the differentiation of peripheral blood pluripotent cells to chondrocytes, comprising the following steps:

[0030] (1) Add sodium citrate anticoagulant to peripheral blood, then add peripheral blood to 0.9% sodium chloride injection, then add 6% hydroxyethyl starch, shake well for 5 minutes, and then let stand 3 hours;

[0031] After layering, absorb the upper cell liquid, and use 4000 rpm to centrifuge for 50 minutes to separate the concentrated solution. The concentrated solution is diluted with normal saline, and then spread on the layered solution prepared by polysucrose and diatrizoate, and then 4000 rpm Centrifuge for 30 minutes, collect the separated stem cell layer, wash the stem cells 3 times with normal saline, and obtain peripheral blood multipotential mesenchymal stromal cells;

[0032] The peripheral blood multipotential mesenchymal stromal cells are taken for primary culture, and the culture medium used in the primary cultu...

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Abstract

The invention relates to a culture method for improving differentiation from peripheral blood multipotent cells to cartilage cells, and belongs to the technical field of cytobiology and regenerative medicine. The culture method comprises the following steps: carrying out primary culture on peripheral blood multipotent mesenchymal matrix cells to obtain third-generation cells; co-culturing the third-generation passed-down peripheral blood multipotent mesenchymal matrix cells and human articular chondrocytes in an induction culture medium. According to the culture method for improving the differentiation from the peripheral blood multipotent cells to the cartilage cells, the peripheral blood multipotent stem cells and the chondrocytes are co-cultured and the two types of cells mutually promote proliferation and the differentiation; the number of times of chondrocyte proliferation and passage and the quantity of the chondrocytes can be reduced, and the differentiation is improved; an efficient method is provided for establishing a seed cell source, and a new medical experiment evidence is also provided for enlarging a cell source of cartilage tissue engineering.

Description

technical field [0001] The invention relates to a culture method for improving the differentiation of peripheral blood pluripotent cells into chondrocytes, belonging to the fields of cell biology technology and regenerative medicine. Background technique [0002] Cartilage tissue is a tissue rich in proteoglycans, collagen, and hyaluronic acid. It is composed of chondrocytes and cartilage matrix. It mainly supports and reduces friction in the body. However, cartilage tissue lacks blood vessels, so it is difficult to repair cartilage tissue once it is damaged, even if it is a small cartilage tissue injury, it is difficult to repair naturally. At present, the clinical methods for repairing cartilage tissue damage mainly include cartilage and subchondral bone removal, microfracture and subchondral bone drilling, autologous osteochondral transplantation, allogeneic osteochondral transplantation, cartilage or periosteum transplantation, etc. These methods can repair cartilage ti...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0655C12N2500/25C12N2500/30C12N2500/32C12N2500/36C12N2500/38C12N2500/80C12N2500/90C12N2501/115C12N2501/15C12N2501/998C12N2502/137C12N2506/1369
Inventor 马亚东黄宗堂曹娜
Owner 丰泽康生物医药(深圳)有限公司
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