Human umbilical cord-derived mesenchymal stem cells and preparation method thereof

A technology of mesenchymal stem cells and human umbilical cord, applied in the field of preparation of primitive mesenchymal stem cells, can solve the problems of long primary cell cycle, decrease in cell number, loss of ability to grow cells, etc., achieve good proliferation potential, and have great application prospects , easy operation effect

Inactive Publication Date: 2020-11-06
国大生命科学产业集团(深圳)有限公司
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  • Claims
  • Application Information

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Problems solved by technology

The disadvantage of this method is that it takes a long time to obtain primary cells, and the buoyancy of the liquid makes the tissue block float up, making it lose the ability to grow cells and reduce the number of cells. In addition, some endothelial cells are mixed in the product, which requires Further screening by post-enzymatic digestion treatment

Method used

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  • Human umbilical cord-derived mesenchymal stem cells and preparation method thereof
  • Human umbilical cord-derived mesenchymal stem cells and preparation method thereof
  • Human umbilical cord-derived mesenchymal stem cells and preparation method thereof

Examples

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Embodiment

[0023] Example: Preparation of human umbilical cord mesenchymal stem cells

[0024] After full-term expectant mothers sign the informed consent form for umbilical cord donation, the mothers are tested for HIV antibody, hepatitis B virus antibody, hepatitis C virus antibody, Treponema pallidum antibody, alanine aminotransferase, mycoplasma, etc., and all test results are negative. Harvest the fetal umbilical cord after delivery.

[0025] 1. After the umbilical cord is removed from the operating table, it is immersed in 0.9% saline containing antibiotics, stored at 4°C and transported to the laboratory;

[0026] 2. After taking out the umbilical cord from the preservation solution in the biological safety cabinet, disinfect it with 75% ethanol and soak it for 5 minutes, wash it three times with PBS containing double antibodies, cut off the part ligated on both sides, and rinse the residual blood on the surface with PBS;

[0027] 3. Remove blood vessels, expose Huatong jelly, an...

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Abstract

The invention discloses a preparation method of human umbilical cord-derived mesenchymal stem cells. According to the preparation method, an umbilical cord of a full-term healthy fetus is adopted to perform aseptic processing, mechanical shearing, exposure of Wharton's jelly, digestion with collagenase, and then separation and culture to prepare the human umbilical cord-derived mesenchymal stem cells, the cell viability does not decrease significantly after cryopreservation and resuscitation, and thereby the human umbilical cord-derived mesenchymal stem cells are an ideal cell source for clinical use. The invention also relates to the mesenchymal stem cells prepared by the method, and the mesenchymal stem cells are confirmed to be mesenchymal stem cells by performing morphology, immunophenotype and in vitro differentiation experiments in accordance with the standards of mesenchymal stem cells formulated by the International Society for Stem Cell Research (ISSCR). The number of cells, cell viability, purity, doubling time, and proliferation ability of the mesenchymal stem cells are significantly higher than those of bone marrow-derived mesenchymal stem cells.

Description

technical field [0001] The invention relates to a preparation method of primitive mesenchymal stem cells, in particular to a preparation method of human umbilical cord-derived mesenchymal stem cells using human umbilical cord Huatong jelly as a material. Background technique [0002] Mesenchymal stem cells (Mesenchymal stem cells, MSCs) are adult stem cells derived from the mesoderm. They are widely found in connective tissues and interstitium of organs throughout the body. They are a type of tissue stem cells with multidirectional differentiation potential. The current clinical trial studies have confirmed that mesenchymal stem cells with self-renewal and multi-directional differentiation potential can be applied to the treatment and rehabilitation of various diseases. However, all research or treatment plans must be based on obtaining a sufficient amount of mesenchymal stem cells. Rapid and effective isolation of mesenchymal stem cells as seed cells has very important rese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2509/00C12N2509/10
Inventor 刘家权牛振宇杨磊
Owner 国大生命科学产业集团(深圳)有限公司
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