Mesenchymal stem cell cryopreservation liquid and activity research method thereof

A technology of mesenchymal stem cells and cryopreservation solution, applied in the medical field, can solve problems such as proliferation, decreased adhesion ability, aging or apoptosis, and difficulty in ensuring ready-to-use

Pending Publication Date: 2021-07-13
HENAN YINFENG BIOENG CO LTD +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] However, excessive passage will show obvious senescence or apoptosis, and long-term in vitro culture is prone to spontaneous differentiation, loss of multi-differentiation potential, decreased proliferation and adhesion, and increased apoptosis rate....

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  • Mesenchymal stem cell cryopreservation liquid and activity research method thereof

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Embodiment Construction

[0017] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0018] see figure 1 , a mesenchymal stem cell cryopreservation liquid and its activity research method, comprising the following steps:

[0019] Step A: Separation of hUC-MSCs, rinse the umbilical cord with sterile saline in an ultra-clean bench, and cut it into smaller than 1mm 3 The tissue pieces were crushed and put into centrifuge tubes, spread the tissue pieces on the bottom of 10 T75 culture flasks, added serum-free medium, and placed in a constant temp...

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Abstract

The invention discloses a mesenchymal stem cell cryopreservation solution and an activity research method thereof. The method comprises the following steps: separating hUC-MSCs, performing in-vitro amplification on the hUC-MSCs, collecting P3 generation cells, performing cooling cryopreservation, performing comparative statistics on the recovered hUC-MSCs and fresh cells, and detecting and identifying the hUC-MSCs. The method has the advantages that a large number of MSCs with high purity are obtained through a serum-free culture adherent method, programmed cooling is conducted through a programmed cooling instrument, damage to cells is reduced, good cell viability is kept, and when the ratio of DMSO to FBS to DMEM is 1: 8.5: 0.5, the survival rate after cryopreservation is optimal, and the morphology and growth rate after recovery are basically consistent with those of normal fresh cell culture.

Description

technical field [0001] The invention relates to a cell activity research method, in particular to a mesenchymal stem cell cryopreservation solution and an activity research method thereof, belonging to the technical field of medicine. Background technique [0002] Mesenchymal stem cells, MSCs are an important member of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development. Mesenchymal stem cells can differentiate into fat, bone, cartilage, muscle, and tendon under specific induction conditions in vivo or in vitro , ligament, nerve, liver, myocardium, endothelium and other tissue cells, which still have multi-directional differentiation potential after continuous subculture and cryopreservation, can be used as ideal seed cells for the repair of tissue and organ damage caused by aging and disease, human umbilical cord Mesenchymal stem cells have become important candidate seed cells for cell therapy, especially in the tissue repair and...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/0775
CPCA01N1/0221A01N1/0284C12N5/0665C12N2509/00C12N2509/10
Inventor 王小龙李雯雯石珂王彦
Owner HENAN YINFENG BIOENG CO LTD
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