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Stem cell preparation for treating knee joint degenerative change and preparation method thereof

A degenerative disease and stem cell preparation technology, applied in the field of stem cells, can solve problems such as large differences between batches, and achieve the effect of avoiding risks, avoiding introduction, and avoiding direct contact

Pending Publication Date: 2022-06-03
深圳汉盛汇融再生医学科技有限公司
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Problems solved by technology

[0007] In order to avoid the uncertainty of the composition of serum in the process of mass cell culture and the problem of large differences between batches, the present invention provides a stem cell preparation for the treatment of knee joint degeneration and its preparation method. The present invention uses DMEM LG / F12 Basal medium, 10% human platelet lysate, 10µg / L basic fibroblast growth factor as culture medium; dexamethasone 10nM, vitamin C 25µg / mL and β-glycerophosphate 2mM as chondrogenic differentiation agent, Successfully achieved the cultivation of umbilical cord mesenchymal stem cells

Method used

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  • Stem cell preparation for treating knee joint degenerative change and preparation method thereof

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Embodiment

A method for preparing stem cells for treating knee joint degeneration, comprising the following steps:

S1. Collection and separation: Take the umbilical cord tissue, cut it into small sections, wash away impurities on the surface of the tissue block, transfer it to a petri dish, dissect out the Wharton's glue and place it in normal saline, and use tissue scissors to separate it into 1- 3mm 3 Tissue blocks, transfer them to culture flasks, add culture medium and disperse them evenly;

S2. Primary culture: place the culture flask at 37°C, 5% CO 2 Cultivate statically in an incubator, change the culture medium every 3 days, and obtain P0 generation umbilical cord mesenchymal stem cells after culturing for 10-20 days; observe the growth of cells with an inverted microscope, and wait until the P0 generation of umbilical cord mesenchymal stem cells grow to cell fusion 70-90%, the cells are in good condition, and the cells are passaged and expanded;

S3. Subculture: Use a pipette...

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Abstract

The invention relates to the technical field of stem cells, in particular to a stem cell preparation for treating knee joint degenerative change and a preparation method thereof. A DMEM LG / F12 basic culture medium, 10% human platelet lysis buffer and 10g / L basic fibroblast growth factors are adopted as culture solutions; 10nM of dexamethasone, 25mu g / mL of vitamin C and 2mM of beta-glycerophosphate are used as chondrogenesis induced differentiation agents, so that the culture of the umbilical cord mesenchymal stem cells is successfully realized. The serum-free culture solution used in the invention can achieve the same culture effect as a culture solution containing serum in a contrast; due to the fact that the used culture solution is clear in component, compared with a culture solution containing fetal calf serum, introduction of animal exogenous substances is avoided, and risks caused by serum to cell culture can be avoided to the maximum extent.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a stem cell preparation for treating knee joint degeneration and a preparation method thereof. Background technique [0002] Knee arthritis is a high-incidence disease among the elderly, with an incidence of 60% over the age of 60, and as high as 80% over the age of 75. With the aging of the social population and people's higher and higher requirements for the quality of life, knee arthritis has become a chronic disease that people are more concerned about, and research on its treatment methods has become an important topic for many medical experts. In recent years, many clinical cases have been obtained by using stem cell regenerative medicine to treat knee arthritis. By inducing stem cells to rapidly differentiate into osteoblasts or chondrocytes, repair damaged tissues, relieve pain, and treat knee arthritis. Good results. [0003] Umbilical cord-derived mesenchymal stem ...

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P19/02
CPCC12N5/0668A61K35/28A61P19/02C12N2509/00C12N2500/90
Inventor 颜浩为李慧娟
Owner 深圳汉盛汇融再生医学科技有限公司
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