Method for promoting multiplication of human adipose-derived stem cells
A technology of human adipose stem cells and cells, which is applied in the field of biological stem cells, can solve the problems that have not yet been seen
Inactive Publication Date: 2018-12-18
淮安康宜莱干细胞生物科技有限公司
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Problems solved by technology
[0006] So far, there is no report that maoshuol, agarwood spirocycline and rohanbocenone can promote the proliferation of adipose-derived stem cells
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[0020] The following describes the substantive content of the present invention in detail in conjunction with the drawings and embodiments, but does not limit the protection scope of the present invention.
[0021] 1. Experimental materials
[0022] Fresh human adipose tissue was obtained from patients undergoing abdominal liposuction, without infectious or endocrine diseases.
[0023] High-glucose DMEM medium and fetal bovine serum were purchased from Gibco, USA. Majool, agarwood spirool and phoenixenone are self-made or purchased, and the purity is greater than 98%.
[0024] Cell counting kit (CCK-8) was purchased from Dojin Chemical Research Institute of Japan Co., Ltd.
[0025] 2. Experimental method
[0026] 1. Isolation and culture of adipose-derived stem cells
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The invention discloses a method for promoting the multiplication of human adipose-derived stem cells. The method comprises the following steps: a fresh human adipose tissue is obtainedtaken, washed with a PBS (phosphate buffer solution), cut into pieces after adipose tissue membranes and blood vessels are removed, and digested, added into a complete medium is added after digestion is completed, undigested fibrous connective tissues are filtered out by a 200-mesh filter screen, centrifuging is carried out, supernate is discarded, suspended adipose cells and lipid droplets are removed, cells are precipitated and resuspended in complete medium, red blood cell lysisate buffer is added, the cells are cultured for lysis under normal temperature after uniform mixing, centrifuging is carried out,and supernate is removed; the cells are precipitated, resuspended in complete medium and inoculated into a culture flask, the culture solution is replaced and the non-adherent cells are removed afterbeing culture for 24 hours of culture, afterwards, solution is replaced every three days, and after being fused, the cells are digested and passage is carried out; the passage cells are extracted anddigested, so that cell suspension is prepared, the cell suspension is inoculated into a culture flask and cultured in complete medium, and after being cultured for 24 hours of culture, the complete medium is replaced by thujopsene-ketone-containing complete medium to continue culture.
Description
technical field [0001] The invention belongs to the field of biological stem cells, and relates to a method for promoting the proliferation of human 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. At present, adipose stem cell-assisted free autologous fat transplantation has been proven to effectively improve the survival rate of transplanted fat. Compared with bone marrow stromal-derived stem cells, adipose-derived stem cells are considered to be the best choice for cell-assisted fat transplantation, because a large number of adipose-derived stem cells can be obtained through liposuction. As a precursor of adipose tissue, adipose-derived stem cells are involved in various adipose tissue remodeling, including growth and developmen...
Claims
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Login to View More IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2500/30C12N2509/00
Inventor 赵立士陈名星
Owner 淮安康宜莱干细胞生物科技有限公司
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