Method for using umbilical stalk placenta to prepare mesenchyma stem cell

A technology of mesenchymal stem cells and placenta, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of streptomycin's potential toxicity to human body and potential danger of treatment, and achieve sensitization and animal source Low content of sexual substances, good proliferation potential, painless effect

Active Publication Date: 2009-07-29
UNION STEMCELL & GENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because penicillin is easy to cause allergic reactions, and streptomycin is potentially toxic to the human body, it is potentially dangerous for mesenchymal cells to be used in the treatment of human diseases in the future

Method used

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  • Method for using umbilical stalk placenta to prepare mesenchyma stem cell

Examples

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Embodiment 1

[0026] Example 1: Preparation method of umbilical cord-derived mesenchymal stem cells

[0027] 1. Take the umbilical cord of a healthy full-term baby and wash the surface with saline solution containing gentamicin (concentration 20,000-40,000 U / L) and heparin (concentration 50-200 U / ml) to remove residual blood and possible bacterial contamination , at least three times, each time the container and equipment are replaced.

[0028] 2. Mechanically crush the tissue, add 4-5 times the volume of 0.01-1% collagenase digestion solution, stir at 37°C, collect the digestion solution after 4-12 hours, filter through a 200-mesh filter, add 2 times the volume of the cell suspension to wash Liquid (DMEM / F12) washed twice, using low fetal bovine serum mesenchymal medium (MesenPRO RS TM Medium, invitrogen) was resuspended and inoculated in a culture bottle, and the medium was changed in full after 24-72 hours, and half of the medium was changed every 3 days.

[0029] 3. When the cells gr...

Embodiment 2

[0038] Example 2: Preparation method of placenta-derived mesenchymal stem cells

[0039] 1. Take the placenta of a healthy full-term baby and wash the surface with saline solution containing gentamicin (20,000-40,000 U / L) and heparin (50-200 U / ml) to remove residual blood and possible bacterial contamination. At least three times, each time the container and instrument are replaced.

[0040] 2. Mechanically crush the tissue, add 4-5 times the volume of collagenase digestion solution, stir at 37°C, collect the digestion solution after 4-12 hours, filter through a 200-mesh filter, add 2 times the volume of washing solution (DMEM / F12) washed twice, using low fetal bovine serum mesenchymal medium (MesenPRORS TM Medium, invitrogen) was resuspended and inoculated in a culture bottle, and the medium was changed in full after 24-72 hours, and half of the medium was changed every 3 days.

[0041] 3. When the cells grow to 80-90% confluence, carry out subculture operation. Discard th...

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Abstract

The invention discloses a method for preparing a mesenchymal stem cell by using umbilical cords and placenta, and mainly relates to a new source for the mesenchymal stem cell and a preparation method thereof. The umbilical cord or the placenta is employed by the stem cell as a cell source; tissues are assimilated at first; then a single cell suspension is prepared and inoculated into a culture bottle, a culture dish or a culture bag to amplify the mesenchymal stem cell, then amplified the mesenchymal stem cell is assimilated, collected and frozen. The mesenchymal stem cell prepared according to the technique of the invention contains a plurality of mesenchymal stem cells with excellent multiplication potentiality. The mesenchymal stem cell can be used in the experimental study, clinical treatment and tissue engineering, etc. The material source thereof is abundant; the separation successful rate and yield are high; the cell multiplication capacity is strong; and the contents of allergic and animal-originated substances are low.

Description

technical field [0001] The invention relates to a method for preparing mesenchymal stem cells, in particular to a method for preparing mesenchymal stem cells by using umbilical cord placenta. Background technique [0002] Mesenchymal stem cells (MSCs) are a research hotspot in recent years. Mesenchymal stem cells have multi-directional differentiation ability, strong in vitro expansion ability, weak antigenicity, and do not involve ethical and moral issues. [0003] Bone marrow mesenchymal stem cells (BMSCs) are undifferentiated cells derived from mesoderm in the bone marrow, which have the potential to differentiate into various tissue cells such as osteoblasts, muscle cells, and vascular endothelial cells, and have high proliferation potential. It has the following advantages: ① It is convenient to obtain materials; ② It is harmless to the health of the donor; ③ It has strong in vitro expansion ability and is not easy to lose the cell phenotype; ④ There is no problem of ti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/0775
Inventor 孟恒星石钏韩俊领黄家学
Owner UNION STEMCELL & GENE ENG
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