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Method and kit for acquiring amniotic mesenchymal stem cells

A technology of amniotic mesenchyme and stem cells, applied in the field of bioengineering, can solve problems that need to be improved

Active Publication Date: 2017-05-10
ACADEMY OF MILITARY MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the current method of obtaining amniotic mesenchymal stem cells still needs to be improved

Method used

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  • Method and kit for acquiring amniotic mesenchymal stem cells
  • Method and kit for acquiring amniotic mesenchymal stem cells
  • Method and kit for acquiring amniotic mesenchymal stem cells

Examples

Experimental program
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Embodiment

[0039] In this example, amniotic mesenchymal stem cells were obtained according to the following method:

[0040] (1) The amniotic membrane stripped from the fresh placenta was cut to a size of 2cm×2cm, washed three times with normal saline, and the blood was washed as much as possible;

[0041] (2) Take 5g of cut amniotic membrane and put it into a 50ml centrifuge tube, add 5mL of 0.25% trypsin-EDTA mixture (containing 0.5mg / mL trypsin and 0.2mg / mL EDTA), at 37°C and 150rpm Shake for 60 minutes, add FBS to stop digestion;

[0042] (3) The digested tissue pieces were washed 3 times with normal saline to minimize the residual trypsin and FBS, then transferred to a 50ml centrifuge tube, added 5mL of 1mg / mL collagenase and 0.2mg / mL DNase I mixture, and incubated at 37°C. Shake for 45 minutes at a temperature of 150 rpm, add 25 ml of normal saline to dilute and terminate, and obtain the digested product;

[0043] (4) Filter the digested product through a 100-mesh sieve, collect ...

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Abstract

The invention discloses a method and a kit for acquiring amniotic mesenchymal stem cells. The method comprises the following steps: carrying out first digestion treatment on an amniotic membrane by adopting a first enzymic preparation, and abandoning digestion liquid, thus obtaining a first digestion product; carrying out second digestion treatment on the first digestion product by adopting a second enzymic preparation, and abandoning digestion liquid, thus obtaining a second digestion product; and separating the amniotic mesenchymal stem cells from the digestion product, wherein the first enzymic preparation comprises a trypsin-EDTA mixed solution, the second enzymic preparation comprises collagenase and DNase I. With the adoption of the method and the kit for acquiring the amniotic mesenchymal stem cells provided by the invention, a number of amniotic mesenchymal stem cells with high purity and high activity can be obtained.

Description

technical field [0001] The invention relates to the field of bioengineering. Specifically, the present invention relates to methods and kits for obtaining amniotic mesenchymal stem cells. Background technique [0002] Placental mesenchymal stem cells are a type of stem cells newly discovered from the placenta in recent years. Compared with stem cells extracted from umbilical cord blood or bone marrow, placental mesenchymal stem cells have a larger number and a wider multidirectional differentiation potential . In addition, the placental tissue comes from postpartum, which is rich in raw materials and convenient to obtain. The collection process does not cause any damage to the mother and newborn, and is not restricted by ethics and clinical medicine. [0003] The placenta consists of three main layers: the amniotic membrane, the chorion, and the decidua basa. The chorion and decidua basalis are rich in blood vessels and red blood cells, and are not easy to separate. The a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2509/00
Inventor 裴雪涛岳文贾雅丽苏如玉
Owner ACADEMY OF MILITARY MEDICAL SCI
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