Method for preparing adipose-derived stem cells by means of extraction

Abstract

The invention discloses a method for preparing adipose-derived stem cells by means of extraction. The method comprises the following steps: acquiring normal abdominal subcutaneous adipose tissue, washing the obtained adipose tissue with a phosphate buffer solution (PBS), and then shearing the adipose tissue into pieces; then, carrying out digestion on the adipose tissue for a period of time by using type I collagenase; after digestion, adding the PBS into the product, evenly mixing and filtering; adding a DMEM culture solution containing human autologous serum, putting into a culture box for culturing, and replacing the culture solution with a fresh culture solution at regular intervals, wherein the cells growing from the tissue are primary cells; when the primary cells reach a certain degree of fusion, washing the cells with the PBS again, and then adding pancreatin and ethylene diamine tetraacetic acid (EDTA) digestive juice; after the cells suspend, adjusting the density of the cells, and adding the DMEM culture solution containing the human autologous serum into the cells again to obtain second-generation cells; and when the second-generation cells reach a certain degree of fusion, adding normal saline into the cells, transferring the cells into a centrifuge tube for carrying out centrifugal filtration, taking supernatant, concentrating and extracting to obtain the adipose-derived stem cells. The adipose-derived stem cells extracted by the method contain multiple life active substances, so that the stem cell properties of the adipose-derived stem cells are further improved.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and relates to a method for extracting and preparing adipose-derived stem cells. Background technique

[0002] Adipose-derived stem cells are a type of stem cells that can self-renew and reproduce, and have the multi-lineage differentiation potential of general stem cells and the stable multi-generational proliferation ability in vitro. It can not only differentiate into adipocytes, osteoblasts, chondrocytes, muscle cells, nerve cells, etc. under the action of different inducing factors, but also secrete a certain amount of cytokines related to cell maturation and various factors that promote angiogenesis and It can act on fibroblasts through paracrine, promote the secretion of type Ⅰ, Ⅲ collagen and fibronectin, promote collagen synthesis, and play anti-inflammatory, anti-oxidation, anti-aging, damage repair and other functions.

[0003] In recent years, the use of living cells for beauty has becom...