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Method for efficiently obtaining adipose-derived mesenchymal stem cells from trace fat

A technique for stem cells and fat cells, which is applied in the field of preparation of adipose-derived mesenchymal stem cells at present, can solve the problems of high requirement of fat sample size and individual limitation of obtaining a large amount of fat, so as to reduce the viscosity of digestion and prevent cell adhesion. Effects of aggregation, superior proliferative activity and differentiation capacity

Pending Publication Date: 2021-09-21
JIANGSU RE STEM BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, the preparation technology of adipose-derived mesenchymal stem cells has become mature, but the requirement for fat sample volume is still relatively high, which is limited for some individuals who are not convenient to obtain a large amount of fat

Method used

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  • Method for efficiently obtaining adipose-derived mesenchymal stem cells from trace fat
  • Method for efficiently obtaining adipose-derived mesenchymal stem cells from trace fat
  • Method for efficiently obtaining adipose-derived mesenchymal stem cells from trace fat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Medium type Final Harvested Cell Volume common complete medium 1.02 billion Complete medium supplemented with substances such as cytokines 2.10 billion

Embodiment 2

[0067] Medium type Final Harvested Cell Volume common complete medium 880 million Complete medium supplemented with substances such as cytokines 1.996 billion

Embodiment 3

[0069] Medium type Final Harvested Cell Volume common complete medium 920 million Complete medium supplemented with substances such as cytokines 2.048 billion

[0070] Second, observe the state of the cells in one of the examples.

[0071] figure 1 It is the morphology of the primary cultured adipose-derived mesenchymal stem cells in the examples under the microscope. figure 2 It is the morphology of the 3rd generation adipose-derived mesenchymal stem cells under the microscope in the example. image 3 It is the morphology of the 5th generation adipose-derived mesenchymal stem cells under the microscope in the embodiment. Depend on Figure 1-Figure 3 It can be seen that the primary culture 9 days after inoculation showed that the cells were uniform in shape, adhered to the wall in a spindle shape, and the cell proliferation ability was active. The cells were distributed in clusters and had good directionality. When the cells are fused to 85%-...

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Abstract

The invention relates to a method for efficiently obtaining adipose-derived mesenchymal stem cells from trace fat. The method comprises the following steps: collecting trace fat; carrying out primary preparation of adipose-derived mesenchymal stem cells; washing the adipose tissue; performing two-step digestion on the adipose tissue by using different digestive enzymes; centrifuging the digested cells; adding a complete culture medium, re-suspending the cell precipitate, and then inoculating the cell precipitate into a coated culture bottle for culturing; carrying out passage on the adipose-derived mesenchymal stem cells; rinsing the cultured fat cells; adding trypsin to digest the adipose cells until the adipose cells are changed into a round wall-off state from a shuttle wall-attaching state under a microscope, and adding DMEM to terminate digestion; transferring the cell suspension into a centrifugal tube for centrifugation; adding cells at the bottom of the centrifugal tube into a complete culture medium, resuspending cell precipitates, and inoculating into a culture bottle for culturing; and subculturing the cells. According to the method, the fat amount is extremely small, and the final yield of cells is not affected.

Description

technical field [0001] The invention relates to the current preparation technology of adipose-derived mesenchymal stem cells, in particular to a method for efficiently obtaining adipose-derived mesenchymal stem cells from a small amount of fat. Background technique [0002] At present, the preparation technology of adipose-derived mesenchymal stem cells has become mature, but the requirement for fat sample volume is still relatively high, which is limited for some individuals who are not convenient to obtain a large amount of fat. [0003] In the prior art, the requirement for fat sample volume generally ranges from 10ml to 300ml. The following documents all mention the separation of adipose-derived mesenchymal stem cells from 10ml-300ml fat. In these documents, there is no clear requirement for the final amount of cells obtained, but only stays at the level of cell acquisition and identification. [0004] Literature [1] Zuk PA, Zhu M, Mizuno H, et al.Multilineage cells fro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2509/00C12N2533/54C12N2533/52C12N2500/90C12N2501/11C12N2501/13C12N2501/105C12N2501/135C12N2501/392C12N2501/999C12N2500/76
Inventor 焦阳张勇王铁朱迪·德瓦库玛杨丽华李靖
Owner JIANGSU RE STEM BIOTECH
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