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Method of separating and cultivating umbilical cord mesenchymal stem cells

A technology of umbilical cord tissue and time, applied in the fields of biomedicine and stem cells, can solve the problems of poor cell viability of UC-MSCs and difficult collection of UC-MSCs, and achieve the effect of reducing the use of enzymes, increasing the amount obtained, and fully digesting

Active Publication Date: 2015-03-25
BGI SHENZHEN CO LTD
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Problems solved by technology

In this way, the method of enzymatic digestion can not only solve the problem of difficult collection of UC-MSCs after Wharton's colloid is digested by collagenase and trypsin, but also solve the problem of Wharton's colloid after being digested by hyaluronidase, collagenase and trypsin for a long time. The problem of poor cell viability of UC-MSCs after co-digestion

Method used

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  • Method of separating and cultivating umbilical cord mesenchymal stem cells
  • Method of separating and cultivating umbilical cord mesenchymal stem cells
  • Method of separating and cultivating umbilical cord mesenchymal stem cells

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Experimental program
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Embodiment

[0059] 1) Under sterile conditions, the umbilical cords of full-term healthy newborns were cut and stored in PBS buffer containing 200U / ml penicillin / streptomycin (Beyond, ST488) at 4°C. The maximum storage time shall not exceed one week.

[0060] 2) In the ultra-clean workbench, cut the umbilical cord of about 3 cm, wash it with PBS buffer, and squeeze out the residual blood clot in the blood vessel.

[0061] 3) Cut the basement membrane of the umbilical cord longitudinally with scissors, and remove 2 arteries and 1 vein in the umbilical cord.

[0062] 4) Wash 3 times with PBS buffer to wash as much blood on the tissue as possible.

[0063] 5) Bluntly separate the Wharton's colloid except the basement membrane of the umbilical cord, collect the colloid and cut it into pieces of about 1mm 3 The size of the organization block.

[0064] 6) Mixture of tissue block and low-sugar DMEM (GIBCO, c11885500bt) containing 0.05% trypsin (GIBCO, 15090-046) and 200U / ml type IV collagenas...

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Abstract

The invention belongs to the biomedical field and particularly belongs to the field of stem cells. The invention provides a method of separating UC-MSCs (umbilical cord mesenchymal stem cells). The method comprises the following steps: 1) mixing umbilical cord tissue pieces with pancreatin and collagenase to incubate; and 2) adding hyaluronidase to incubate.

Description

technical field [0001] The invention belongs to the field of biomedicine, more specifically to the field of stem cells. Background technique [0002] Stem cells are a kind of cells capable of self-renewal and multi-directional differentiation potential, and have broad application prospects in tissue engineering and cell therapy. Stem cells can be divided into two categories: embryonic stem cells and adult stem cells. [0003] Embryonic stem cells (Embryonic Stem Cells, ESCs) are a type of highly undifferentiated cells with totipotency. However, the source of ESCs is difficult, and it is easy to cause safety and ethical issues, which greatly limits the research and application of ESCs. [0004] Adult stem cells (Adult Stem Cells) are a type of pluripotent cells that exist in differentiated tissues, which are capable of self-renewal and can specialize to form cells that make up this type of tissue. Mesenchymal Stem Cells (MSCs) are a type of adult stem cells derived from th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 吴亮商周春刘聪王全磊陈杰董国艺葛玉萍李静
Owner BGI SHENZHEN CO LTD
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