A preparation method of high-concentration human-derived mesenchymal stem cell lysate

Abstract

The invention provides a method for preparing a high-concentration human-derived mesenchymal stem cell lysate, comprising the following steps: crushing step: using 60-100W ultrasonic waves to collect human umbilical cord mesenchymal stem cells with a fusion degree of 80-90% Ultrasonic crushing for 10-30min to obtain a broken cell solution; centrifugation step: the broken cell solution is centrifuged for 10-30min at a temperature of 2-8°C and a centrifugal force of 2000-5000g, and the supernatant is taken after centrifugation; purification steps: Utilize 3K ultrafiltration membrane to filter and concentrate the supernatant to obtain a concentrated solution with a volume of 1 / 5-1 / 10 of the volume of the supernatant; utilize a hollow fiber ultrafiltration column to further process the concentrated solution Concentrate to obtain a high-concentration human-derived mesenchymal stem cell lysate. The method of the present invention overcomes the problems of low purity, low yield, low activity, low efficiency, and low yield of extracted stem cell bioactive factors in the prior art; the obtained stem cell lysate has a good ability to promote the proliferation of mesenchymal stem cells and promote Ability to synthesize collagen.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing a high-concentration human-derived mesenchymal stem cell lysate. Background technique [0002] Stem cells, as emerging seed cells, are cells with multi-directional differentiation potential. Transplanted stem cells can self-renew and differentiate into various cells to repair tissues; and stem cells can produce or secrete a large number of biologically active factors. These biologically active Functionally, the factors can be divided into six categories: immune regulation, anti-apoptosis, angiogenesis, support for proliferation and differentiation of stem / progenitor cells, chemotaxis, and anti-scarring. Through autocrine or paracrine methods, they improve the local microenvironment, promote the migration and differentiation of endogenous stem cells to the wound site, and then physiologically repair or replace the damaged, diseased and aging cells of the body. ...

AI Technical Summary

Problems solved by technology

At present, the method for extracting stem cell bioactive factors in practice has the following deficiencies: (1) direct use of tissues such as placenta to extract, the obtained active protein contains a large amount of tissue protein that is difficult to remove, and the obtained stem cell bioactive factors have low purity; (2) Utilizing stem cell expansion culture fluid to extract active factors requires concentrating a large amount of culture fluid, which is time-consuming and laborious, and the concentration of active factors contained in the culture fluid is relatively low, that is, the extraction efficiency is low and the yield is low; (3) stem cells contain abundant The biologically active substances of stem cells play an important role in the proliferation and differentiation of stem cells, but such substances are not secreted outside the cells during the process of proliferation and differentiation