Stem cell culture medium as well as preparation method and application thereof

A stem cell and culture medium technology, applied in the field of stem cell culture medium and its preparation, can solve the problems of inability to adapt to menstrual blood stem cells, poor cell quality, low proliferation efficiency, etc., to improve the purity and proliferation ability, promote cell growth, and strongly promote mitosis. Effect

Inactive Publication Date: 2018-06-29
GUANGZHOU ZISHENG BIOLOGICAL TECH CO LTD
View PDF12 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current culture system cannot adapt to menstrual blood stem cells. After using the existing stem cell culture medium, the proliferation efficiency of menstrual blood stem cells is low, and the cell quality is poor; Meet the demand of clinical cell dosage
[0005] In summary, traditional stem cell culture medium is not suitable for the culture of menstrual blood stem cells

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Stem cell culture medium as well as preparation method and application thereof
  • Stem cell culture medium as well as preparation method and application thereof
  • Stem cell culture medium as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] This example is the separation of menstrual blood stem cells. References for the separation method: Wang Xueli, Experimental Research on the Biological Characteristics and Multidirectional Differentiation Potential of Menstrual Blood-derived Endometrial Stem Cells, Master's Degree Thesis of Anhui Medical University [M], 2016.

Embodiment 2

[0032] This embodiment is the cultivation of menstrual blood stem cells, and the cultivation steps are as follows:

[0033] a. Add 0.01% poly-lysine to the culture dish, make the liquid soak the bottom of the dish, and incubate at 37°C, and discard the poly-lysine after 1 hour.

[0034] b. Configure the stem cell culture medium of components 1 to 5 according to Table 1.

[0035] c. Resuspend menstrual blood stem cells with the stem cell culture medium of components 1 to 5, and adjust the density to 8×10 4 ~1×10 5 cells / mL, inoculated into poly-lysine-incubated petri dishes.

[0036] d. Transfer cells to 37°C, 5% CO 2 , 95% humidity environment, and change the medium after 48 hours.

[0037] e. When the cell fusion rate reaches about 80%, carry out cell subculture.

[0038] Table 1 Composition Table of Group 1 to Group 5

[0039]

Embodiment 3

[0041] This embodiment is the surface antigen detection of menstrual blood stem cells, and the specific experimental steps are as follows:

[0042] The menstrual blood stem cells cultured in Group 1 to Group 5 in Example 2 were taken to make a cell suspension of 1×106, and anti-human CD34, CD45, CD19, CD73, CD90, CD105, CD11b, and HLA-DR were taken respectively. Add 5 μL of each cloned antibody to 500 μL of cell suspension, incubate at room temperature in the dark for 20 min, set up a blank isotype control, centrifuge at 1500 r / min for 5 min, discard the supernatant, wash twice with PBS containing 10% FBS, and resuspend in 500 μL PBS Then check on the machine.

[0043]

[0044] The expression rates of CD73, CD90, and CD105 of mesenchymal stem cells are above 95.0%, and the expression rates of CD34, CD45, CD19, CD11b, and HLA-DR are below 2.0%. The menstrual blood stem cells obtained in the present invention meet the identification standard of mesenchymal stem cells,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention belongs to the technical field of cell culture, and in particular relates to a stem cell culture medium as well as a preparation method and application thereof. The stem cell culture medium provided by the invention comprises a mesenchymal stem cell serum-free medium, platelet lysate, glutamine, non-essential amino acids and a hepatocyte growth factor, wherein the mesenchymal stem cell serum-free medium and platelet lysate enable menstrual blood stem cells to rapidly grow, the glutamine and the non-essential amino acids provide the nutrients needed by the menstrual blood stem cells, and the hepatocyte growth factor stimulates the proliferation of the menstrual blood stem cells. The stem cell culture medium solves the technical problem that the stem cell culture medium in theprior art is not suitable for culturing the menstrual blood stem cells.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a stem cell culture medium and its preparation method and application. This application claims the rights and interests of the Chinese patent (patent application number 201810065267.7) submitted on January 23, 2018, and the entire content of the above application is hereby incorporated by reference. Background technique [0002] Stem cells refer to a class of early undifferentiated cells with high proliferation potential, self-renewal ability and high differentiation ability. Mesenchymal stem cells are currently the most concerned member of stem cells, with high self-renewal ability and multi-directional differentiation potential. The human uterus is a tissue with a high ability to regenerate and differentiate. The endometrium contains stem cells with a high ability to proliferate. Scientists can separate these stem cells from the menstrual blood of healthy wome...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N2501/12C12N2500/32C12N2500/80C12N2500/90
Inventor 马颖马铎
Owner GUANGZHOU ZISHENG BIOLOGICAL TECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap