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

Umbilical cord tissue digestion method

A technology of human umbilical cord and tissue, which is applied in the field of cell separation, can solve the problems of destroying cell surface proteins, single types of digestive enzymes, and inconsistent selection of enzymes and digestion time. The effect of complete separation

Inactive Publication Date: 2017-08-15
青岛青春派生物科技有限公司
View PDF6 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Enzyme digestion method has no uniform standard for enzyme selection and digestion time, and the existing problems mainly include: it takes a long time to completely digest tissue blocks; there are only one type of digestive enzymes, which cannot completely digest hyaluronic acid and collagen fibers; simply increasing the dosage of enzymes, It will destroy the free cell surface proteins such as adhesion factors, which will affect the cell viability and adhesion ability

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
  • Umbilical cord tissue digestion method
  • Umbilical cord tissue digestion method
  • Umbilical cord tissue digestion method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A human umbilical cord tissue digestion method, the method steps are as follows:

[0029] Preparation of tissue-digesting enzymes: put collagenase, hyaluronidase and DNase into DMEM medium, filter and sterilize through a 0.22um filter membrane, and pre-warm at 37°C for later use; collagenase, hyaluronidase and DNA The mass ratio of enzyme added was 40:40:1.

[0030] S1: Umbilical cord pretreatment: In the delivery room, remove the umbilical cord from the fetus and put it in the umbilical cord transport solution, and refrigerate it at 4°C for later use; take out the umbilical cord tissue and wash it twice in the pre-cooled tissue cleaning solution; cut the cleaned umbilical cord tissue into 4- 5cm section, peel off the outer wall of the umbilical cord and blood vessels in the cleaning solution to obtain a transparent jelly-like substance that is Wharton's jelly;

[0031] S2: Primary digestion: Wash the Wharton's jelly obtained in S1 and cut it into 1-2mm pieces 3 Add 1...

Embodiment 2

[0036] A human umbilical cord tissue digestion method, the method steps are as follows:

[0037] Preparation of tissue-digesting enzymes: put collagenase, hyaluronidase and DNase into DMEM medium, filter and sterilize through a 0.22um filter membrane, and pre-warm at 37°C for later use; collagenase, hyaluronidase and DNA The mass ratio of enzyme addition is 60:60:1.

[0038] S1: Umbilical cord pretreatment: In the delivery room, remove the umbilical cord from the fetus and put it in the umbilical cord transport solution, and refrigerate it at 4°C for later use; take out the umbilical cord tissue and wash it twice in the pre-cooled tissue cleaning solution; cut the cleaned umbilical cord tissue into 4- 5cm section, peel off the outer wall of the umbilical cord and blood vessels in the cleaning solution to obtain a transparent jelly-like substance that is Wharton's jelly;

[0039] S2: Primary digestion: Wash the Wharton's jelly obtained in S1 and cut it into 1-2mm pieces 3 Add...

Embodiment 3

[0044] A human umbilical cord tissue digestion method, the method steps are as follows:

[0045]Preparation of tissue-digesting enzymes: put collagenase, hyaluronidase and DNase into DMEM medium, filter and sterilize through a 0.22um filter membrane, and pre-warm at 37°C for later use; collagenase, hyaluronidase and DNA The mass ratio of enzyme addition is 50:50:1.

[0046] S1: Umbilical cord pretreatment: In the delivery room, remove the umbilical cord from the fetus and put it in the umbilical cord transport solution, and refrigerate it at 4°C for later use; take out the umbilical cord tissue and wash it twice in the pre-cooled tissue cleaning solution; cut the cleaned umbilical cord tissue into 4- 5cm section, peel off the outer wall of the umbilical cord and blood vessels in the cleaning solution to obtain a transparent jelly-like substance that is Wharton's jelly;

[0047] S2: Primary digestion: Wash the Wharton's jelly obtained in S1 and cut it into 1-2mm pieces 3 Add ...

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 discloses an umbilical cord tissue digestion method. According to the invention, wharton jelly is obtained from umbilical cord, twice digestion is carried out by using different tissue digestive enzyme, filtering is carried out, and the required mesenchymal stem cells can be obtained through filtering and centrifugation. The method employs twice digestion, during primary digestion, collagenase, hyaluronidase and DNA enzyme are subjected to complex formulation for usage, wharton jelly is digested, the digestive enzyme can effectively reduce the digestion consistency and has small damage on cells; pancreatin is employed during secondary digestion, and the cell separating can be more completed. The provided method can avoid the influence of long-term digestion of digestive enzyme such as collagenase and pancreatin on cell bodies, the acquisition rate of the obtained mesenchymal stem cells is high, and the obtained mesenchymal stem cells have high activity and differentiation potential.

Description

technical field [0001] The invention relates to the technical field of cell separation, in particular to a method for digesting human umbilical cord tissue. Background technique [0002] Mesenchymal stem cells (MSCs) can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells under specific induction conditions in vivo or in vitro, and After continuous subculture and cryopreservation, it still has multi-directional differentiation potential, and can be used as an ideal seed cell for the repair of tissue and organ damage caused by aging and disease. Mesenchymal stem cells mainly include bone marrow mesenchymal stem cells, fat mesenchymal stem cells and umbilical cord mesenchymal stem cells according to the tissue barrier. Among them, compared with bone marrow mesenchymal stem cells, umbilical cord mesenchymal stem cells have the advantages of abundant sources, low cost, and no damage to donors, and h...

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
CPCC12N5/0668C12N2509/00
Inventor 沈晓延
Owner 青岛青春派生物科技有限公司
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