Separation and culture of mouse mesenchymal stem cells

A technology of bone marrow mesenchymal and stem cells, applied in the field of cell culture of modern biotechnology, can solve the problems of difficult operation and high cost, influence of cell activity, insufficient cell purity, etc., and achieve easier storage, high cell purity, and large number of cells Effect

Inactive Publication Date: 2019-10-29
JIANGYIN CHI SCI
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The whole bone marrow method, also called the adherent method, is the earliest method for the isolation of bone marrow mesenchymal stem cells. The whole bone marrow method is a culture method in which all the flushing fluid washed out is centrifuged and washed directly, and then inoculated directly. Blood cells and other non-adherent cells are removed by cha

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
  • Separation and culture of mouse mesenchymal stem cells
  • Separation and culture of mouse mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0021] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0022] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0023] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0024] The experimental instruments and reagents used in this experiment are as follows: a set of surgical instruments, CCK-8 cell counting box (Sigma, USA), inverted microscope (XDS-1A, Shanghai), fluorescence microscope (Leica, USA), cryogenic centrifuge ( TD24B-WS, Shanghai), ultra-low temperature refrigerator (Zhongke Meiling), pipette gun (Eppendorf, USA), electronic analytical balance (Sartorius, USA), ultra-clean bench (HJ-CJ-1D, Shanghai), CO 2 Cell culture box (SA...

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 provides a separation and culture method of mouse mesenchymal stem cells. The separation and culture method comprises the following steps: (1) an ICR mouse is anesthetized and killed; (2) thighbones and tibiae are taken and placed in ethyl alcohol of 75% to be soaked and disinfected; (3) washing is conducted for three times through PBS, and the epiphysis ends of the thighbones and the tibiae are cut off; (4) a culture medium containing double resistants is sucked through a needle tube of 10 ml to rush out bone marrow; (5) a percoll separating medium is used for centrifuging through a density gradient method, and a middle white layer is taken; (6) washing is conducted for two times through the PBS; (7) the cell activity is detected; (8) the cells are counted; (9) the cell morphology is identified; (10) detecting is conducted through an ELISA method; and (11) RT-PCR detection is conducted. The separation and culture method of the mouse mesenchymal stem cells is easy to operate, large in number of the obtained cells, high in survival rate and ideal, and provides the reliable cell resource for experiments.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, and specifically relates to a method for separating and culturing mouse bone marrow mesenchymal stem cells. . Background technique [0002] Bone marrow mesenchymal stem cells are important members of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development. Bone marrow mesenchymal stem cells were originally discovered in the bone marrow, and have attracted increasing attention because of their multi-lineage differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. For example, mesenchymal stem cells can be differentiated into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, heart muscle, endothelial and other tissue cells under specific induction conditions in vivo or in vitro, after continuous subculture and cryopreservation It still has multi...

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
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2501/33C12N2533/32
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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