Method for extracting paracrine factor from adipose-derived stem cells

Abstract

The invention discloses a method for extracting a paracrine factor from adipose-derived stem cells. The method comprises the following steps: taking a collected fat sample, and washing the fat samplewith normal saline for 1-3 times; adding a proper amount of adipose cell digestive juice for digestion; performing centrifugation, resuspending cell precipitate, and performing primary culture with aserum-free medium; performing subculture; performing enlarged culture; and performing extraction with a paracrine factor extracting solution, and performing purification to obtain the paracrine factor, wherein the paracrine factor extracting solution is prepared from a PBS buffer solution, L-glutamine, D-glucose and L-ascorbic acid, and the concentrations of the L-glutamine, the D-glucose and theL-ascorbic acid are 1-3mmol / L, 10-20 micromoles / L and 10-100 micromoles / L respectively. The method disclosed by the invention can stably extract the adipose-derived stem cells from adipose tissues, and extract the paracrine factor of the adipose-derived stem cells by utilizing the adipose-derived stem cells, so that the cell quality and the factor extraction efficiency are stable; and the adipose-derived stem cells are separated and cultured for a long time in a GMP environment, do not contact any animal-derived component, and can efficiently express adipose-derived stem cell markers such as CD29, CD44 and CD105.

Description

technical field [0001] The invention relates to a method for extracting paracrine factors from fat stem cells, belonging to the technical field of cell biology. Background technique [0002] Paracrine means that the producing cell of a certain cytokine and the target cell are not the same cell, but they are adjacent; the biological effect of the factor on the target cell is the paracrine effect. Paracrine signaling molecules include growth factors, lymphokines, etc. Current studies have found that stem cells from a variety of tissue sources can secrete and produce different cytokines, that is, the paracrine effect of stem cells is universal. [0003] Mesenchymal stem cells have been used in the treatment of more than ten kinds of refractory diseases. In addition to promoting the recovery of hematopoiesis and co-transplantation with hematopoietic stem cells to improve leukemia and refractory anemia, etc., they are also used for cardiovascular and cerebrovascular diseases, li...

AI Technical Summary

Problems solved by technology

[0005] At present, the research on stem cell paracrine factors in the industry has just started, and most of them are mainly in the research stage, without transformation into the production stage, and there are no clear industry standards and production quality standards. There are the following shortcomings: the source of paracrine factor samples is not standardized, The culture scale is small, the quality levels are uneven, the process of inducing cytokine secretion is not efficient enough, the separation and purification process is not scaled, and the product shelf life is short, which greatly limits the production and use of paracrine factors

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