Preparation method of human umbilical cord mesenchymal stem cell factor freeze-dried powder

Abstract

The invention relates to a preparation method of human umbilical cord mesenchymal stem cell factor freeze-dried powder, and aims to solve the problems of low cell factor yield and poor activity in the prior art. Umbilical cord mesenchymal stem cells are sequentially separated and subjected to passage, culture supernatant fluid is collected, filtered and concentrated, freeze-dried excipients and 50 DEG C deionized water are added, protein concentration is adjusted, and the materials are blended, sub-packaged and then freeze-dried under the freeze-drying condition of 50-100Pa pressure intensity and temperature ranging from -30 DEG C to -35 DEG C to obtain the human umbilical cord mesenchymal stem cell freeze-dried powder. The collected culture supernatant fluid is firstly frozen and stored, filtering and concentrating processes of the culture supernatant fluid include unfreezing the culture supernatant fluid at room temperature and filtering the cells by 0.45 micrometer filter membranes, ultra-filtration and concentration of 3KD filter membranes include concentrating the supernatant fluid to be 1/10 of the original volume of the supernatant fluid, and 0.22 micrometer filter membranes perform filtration sterilization to obtain the human umbilical cord mesenchymal stem cell freeze-dried powder stock solution for preparing the human umbilical cord mesenchymal stem cell freeze-dried powder. The prepared freeze-dried powder can effectively store various cell factors secreted by the human umbilical cord mesenchymal stem cell and with biological activity, and the obtained cell factors are high in yield, good in activity and easy to store and transport.

Description

technical field [0001] The invention relates to a preparation method of freeze-dried powder, in particular to a preparation method of human umbilical cord mesenchymal stem cell factor freeze-dried powder. Background technique [0002] Umbilical cord mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) refer to a kind of multifunctional stem cells existing in the umbilical cord tissue of newborns, which can differentiate into many kinds of tissue cells and have broad clinical application prospects. The cell content and proliferation ability are better than those of bone marrow MSCs, and the immunogenicity is lower than that of bone marrow MSCs, and it has the advantages of convenient sampling and no ethical disputes. [0003] Studies have shown that umbilical cord mesenchymal stem cells can secrete a variety of cytokines during the culture process, such as secreting epidermal growth factor (epidermal growth factor, EGF), hepatocyte growth factor (hepatocyte growth factor, HG...

AI Technical Summary

Problems solved by technology

[0004] In recent years, with the continuous deepening of stem cell research, the application of stem cell products to the beauty industry has attracted more and more attention. Most of the cyt