Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for carrying out in-vitro large-scale culture on umbilical cord mesenchymal stem cells

A technology of quality stem cells and umbilical cords, applied in the field of stem cell culture, to achieve the effects of improving adsorption and growth speed, ensuring biogenicity and safety, and increasing yield

Inactive Publication Date: 2017-02-22
CHONGQING STEM CELL TECH +7
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are certain controversies about the safety of mesenchymal stem cells, so whether umbilical cord mesenchymal stem cells are tumorigenic is also a hot spot in current research

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
  • Method for carrying out in-vitro large-scale culture on umbilical cord mesenchymal stem cells
  • Method for carrying out in-vitro large-scale culture on umbilical cord mesenchymal stem cells
  • Method for carrying out in-vitro large-scale culture on umbilical cord mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Collect the umbilical cord and cord blood of full-term infants according to national standards, and disinfect the umbilical cord with alcohol;

[0040] (2) Wash off most of the residual blood with fresh buffer solution (containing 1.00 v / v% penicillin / streptomycin), and retain more than 10 v / v% of residual uncoagulated blood. The umbilical cord was crushed using an instrument to obtain 1mm 3 Large and small tissue fragments are digested for a sufficient time in a low-concentration collagenase II environment;

[0041] (3) Discard the remaining tissue pieces after digestion, add eluent and use four-step centrifugation to separate unpurified original umbilical cord mesenchymal stem cells. The parameters of centrifugation are: 300r / min for 10 minutes, 1800r / min for 15 minutes, and 1000r / min10 minutes, 1000r / min10 minutes;

[0042] (4) Count the obtained primary umbilical cord mesenchymal stem cells according to 5.0×10 4 / cm 2 ~1.0×10 5 / cm 2 Planted in a growing ...

Embodiment 2

[0050] Embodiment 2, step (6) in embodiment 1 is pressed 1.0 * 10 7 The concentration per bottle was inoculated in the HYPERFlask culture vessel as compared to 1.25×10 7 / bottle, 1.0×10 7 / bottle, 0.75×10 7 / bottle, 0.5×10 7 The concentration per bottle was inoculated in the HYPERFlask culture vessel.

Embodiment 3

[0051] In Example 3, step (6) in Example 1 was inoculated into a HYPERFlask culture vessel as opposed to being inoculated into a HYPERFlask culture vessel and a common cell culture vessel.

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 relates to a method for carrying out in-vitro large-scale culture on umbilical cord mesenchymal stem cells. The method comprises the steps of: cleaning away most of the residual blood of an umbilical cord; preparing tissue fragments from the umbilical cord, and carrying out digestion by collagenase II; removing the tissue fragments after digestion, adding eluant to carry out centrifugation, and separating out unpurified first-generation umbilical cord mesenchymal stem cells; inoculating the obtained first-generation stem cells to a culture vessel according to 5.0*104 / cm<2> to 1.0*105 / cm<2>; after basal layer adherent cells are fused by 80% or more, carrying out digestion, and carrying out passage inoculation on the digested and dispersed cells to the culture vessel until obtaining cells subjected to four times of passage; inoculating fourth-generation cells to a HYPERFlask culture vessel, and until obtaining fifth-generation cells or sixth-generation cells, ending culture. According to the method disclosed by the invention, the first-generation umbilical cord mesenchymal stem cells are sufficiently subjected to in-vitro high-efficiency amplification by utilizing the HYPERFlask cell culture vessel, and a great quantity of mesenchymal stem cells applied to clinic and research are obtained.

Description

technical field [0001] The invention relates to a method for culturing stem cells, in particular to a method for culturing umbilical cord mesenchymal stem cells, so as to realize large-scale culture in vitro. Background technique [0002] According to the clinical trial registration system (Clinicaltrials.gov) managed by the National Institutes of Health in the United States, as of May 2016, there were 612 registered mesenchymal stem cell clinical research projects worldwide, including 125 in China. Analysis of 567 clinical trials of registered MSCs in the world, in addition to promoting recovery of hematopoiesis, co-transplantation with hematopoietic stem cells to improve leukemia and refractory anemia, etc., it is also used for cardiovascular and cerebrovascular diseases, liver cirrhosis, bone and muscle degenerative diseases , Brain and spinal cord injury, senile dementia, and autoimmune diseases such as lupus erythematosus and scleroderma. The results of some clinical tr...

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
CPCC12N5/0668
Inventor 章毅陈亮王莎萍王波
Owner CHONGQING STEM CELL TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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