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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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,...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com