Preparation method of high-concentration human-source mesenchymal stem cell lysis solution

Abstract

The invention provides a preparation method of a high-concentration human-source mesenchymal stem cell lysis solution. The preparation method comprises the following steps of step of breaking: performing ultrasonic breaking on a human-source mesenchymal stem cell lysis solution of which the fusion degree is 80-90%, which is obtained by collection by using 60-100W ultrasonic waves for 10-30min to obtain broken cell liquid; step of centrifugation: performing centrifugation on the broken cell liquid under the condition that the temperature is 2-8 DEG C and centrifugal force is 2000-5000g for 10-30min, and after centrifugation, taking supernatant; step of purification: performing filtering and concentrating on the supernatant by a 3K ultra filtration membrane to obtain a concentrated solutionof which the volume is 1 / 5-1 / 10 of the volume of the supernatant; and performing further concentrating on the concentrated solution by a hollow fiber ultrafiltration column to obtain the high-concentration human-source mesenchymal stem cell lysis solution. The method solves the problems that in the prior art, the extracted stem cell factors are low in purity, low in yield, low in activity, low inefficiency, low in yield and the like, and the obtained stem cell lysis solution has favorable capacity for promoting proliferation of mesenchymal stem cells and promoting collagen synthesis.

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