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A method for promoting the proliferation of fat stem cells

An adipose stem cell and culture medium technology, which is applied in the biological field, can solve the problems that cells cannot survive and proliferate well, and achieve the effect of high proliferation rate

Active Publication Date: 2018-11-27
江西汉氏联合干细胞科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in most cases, the transplanted cells cannot survive and proliferate well, therefore, finding suitable drugs to promote the proliferation of transplanted stem cells is very important to improve the effect of stem cell transplantation treatment

Method used

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  • A method for promoting the proliferation of fat stem cells
  • A method for promoting the proliferation of fat stem cells

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Embodiment Construction

[0016] The technical solutions of the present invention are described below in combination with specific embodiments. The experimental materials that are not particularly emphasized in the following examples are conventional experimental materials without special requirements, and are all conventional materials that are easily obtained by those skilled in the art.

[0017] 1. Experimental materials

[0018] Saccharin A, Saccharin B, Saccharin F and Saccharin F were self-made according to the literature method, and the purity was greater than 98%.

[0019] DMEM medium and fetal bovine serum were purchased from Gibco. CCK-8 kit was purchased from colleagues in Japan.

[0020] 2. Experimental method

[0021] 1. Isolation and culture of adipose-derived stem cells

[0022] The subcutaneous adipose tissue of healthy volunteers was taken, washed three times with PBS buffer, and the adipose tissue membrane and blood vessels were removed with ophthalmic scissors and ophthalmic forc...

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Abstract

The invention discloses a method for facilitating proliferation of adipose-derived stem cells. The method comprises the following steps: (1) taking a subcutaneous fatty tissue, washing the subcutaneous fatty tissue with a PBS buffer solution, removing an adipose tissue membrane and blood vessels and shearing off the subcutaneous fatty tissue into fragments with the sizes being 1 mm<3>, carrying out enzymic digestion, adding a complete medium after digestion is finished, centrifuging, removing supernatant, resuspending cell precipitates on the complete medium, inoculating the cell precipitatesin a culture bottle, changing a culture solution after culturing for 24 h and removing cells which are not attached to walls, then changing the solution every 3 days, digesting for 3-5 min with 0.25%of trypsin and 1 mmol / L EDTA at the temperature of 37 DEG C after cell fusion, stopping digestion of the complete medium, and carrying out passage at the ratio of 1: 2; and (2) inoculating passage cells into a culture bottle which contains the complete medium at the density of 1.5* 104 / cm<2>, culturing for 48 h and then changing the culture medium into an incomplete medium, meanwhile adding a jatrophane type diterpenoid compound and continuing culturing for 24-72 h. Surface euphorbia helioscopia terpene A, surface euphorbia helioscopia terpene B and surface euphorbia helioscopia terpene F canbe used as an accelerant for proliferation of adipose-derived stem cells, and can be added in a DMEM medium to obtain an adipose-derived stem cell high-proliferation medium.

Description

technical field [0001] The invention belongs to the biological field and relates to a method for promoting the proliferation of fat stem cells. Background technique [0002] Adipose stem cells are a type of pluripotent stem cells in adipose tissue, which can differentiate into various cell types under specific conditions in vivo and in vitro, such as adipocytes, osteoblasts, chondrocytes, nerve cells, muscle cells, etc. Adipose stem cells can be used as an ideal cell source for tissue regeneration engineering because they are easy to isolate, can be expanded in large quantities under in vitro conditions and can be stably cultured for a long time. In the process of cell transplantation tissue reconstruction, it is necessary to provide a sufficient source of cells for the formation of new tissue. However, in most cases, the transplanted cells cannot survive and proliferate well. Therefore, it is very important to find suitable drugs to promote the proliferation of transplante...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2501/999C12N2509/00
Inventor 韩之波田红青
Owner 江西汉氏联合干细胞科技有限公司
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