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Preparation and storage of umbilical cord-mesenchymal stem cells (UC-MSCs) used in clinical therapy

A stem cell and umbilical cord technology, which is applied in the field of preparation and storage of human umbilical cord mesenchymal stem cells, can solve the problems of increasing the safety risk of clinical application, existing allergies and cross-infection, and low purity of primary cultured cells, and achieves good application prospects. , excellent effect, good safety effect

Inactive Publication Date: 2009-12-02
THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the enzymatic hydrolysis method requires the application of animal protein-derived digestive enzymes, there is a possibility of allergy and cross-infection, which increases the safety risk of clinical application
In addition, the cost of enzymatic hydrolysis is high, and the purity of primary cultured cells is low

Method used

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  • Preparation and storage of umbilical cord-mesenchymal stem cells (UC-MSCs) used in clinical therapy
  • Preparation and storage of umbilical cord-mesenchymal stem cells (UC-MSCs) used in clinical therapy

Examples

Experimental program
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Effect test

Embodiment 1

[0021] 1. Under aseptic conditions, take the umbilical cord of the normal delivery fetus, the umbilical cord is about 20cm long, and wash it repeatedly with normal saline containing 2% gentamicin to remove the residual blood. At the same time, the pathogenic microorganism detection is carried out, and the following steps are carried out after being completely qualified.

[0022] 2. Place the umbilical cord in physiological saline containing 2% gentamicin, cut it into particles with sterile scissors, collect the tissue pieces and place them in a 50ml centrifuge tube, and centrifuge at a high speed at 2000 rpm , centrifuged for 10 minutes, discarded the supernatant, re-added physiological saline containing 2% gentamicin, fully oscillated the precipitated tissue pieces to make them loose, and centrifuged at a high speed at 2000 rpm for 10 minutes. .

[0023] 3. Suspend the tissue block obtained in step 2 in DMEM medium containing 10% fetal bovine serum, shake fully and centrifug...

Embodiment 2

[0031] 1. Under aseptic conditions, take the umbilical cord of the fetus delivered by cesarean section, the umbilical cord is about 25cm long, and wash it repeatedly with physiological saline containing 2% gentamicin to remove residual blood. At the same time, the pathogenic microorganism detection is carried out, and the following steps are carried out after being completely qualified.

[0032] 2. Place the umbilical cord in physiological saline containing 2% gentamicin, cut it into particles with sterile scissors, collect the tissue pieces and place them in a 50ml centrifuge tube, and centrifuge at a high speed at 2000 rpm , centrifuged for 10 minutes, discarded the supernatant, re-added physiological saline containing 2% gentamicin, fully oscillated the precipitated tissue pieces to make them loose, and centrifuged at a high speed at 2000 rpm for 10 minutes. .

[0033] 3. Suspend the tissue block obtained in step 2 in DMEM medium containing 10% fetal bovine serum, shake fu...

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Abstract

The invention relates to preparation and storage of umbilical cord-mesenchymal stem cells (UC-MSCs) used in clinical therapy. The preparation method of the UC-MSCs comprises the following steps: cleaning an umbilical cord and cutting the umbilical cord into granules; putting the granules in physiological saline containing 2 percent gentamicin; carrying out centrifugation, suspension and centrifugation; carrying out cultivation for 10 days; carrying out passage in the proportion of 1:3 to between fourth and fifth generation, and obtaining a great amount of mesenchymal stem cells; and finally, putting the mesenchymal stem cells in liquid nitrogen for storage after adding freezing medium and keeping the freezing temperature of the liquid nitrogen at 196 DEG C below zero.

Description

technical field [0001] The invention relates to the preparation of a medical product, in particular to the preparation and storage of human umbilical cord mesenchymal stem cells for clinical treatment. technical background [0002] Mesenchymal stem cells (MSCs) are a type of stem cells named for their ability to differentiate into mesenchymal tissues. Potential for tissue differentiation and development. MSCs mainly exist in connective tissue and organ interstitium, and are most abundant in bone marrow. They can differentiate into tissue cells derived from a variety of mesoderm and neuroectoderm, such as osteoblasts, chondrocytes, adipocytes, and tendons. Cells, smooth muscle cells, bone marrow stromal cells, fibroblasts and various vascular endothelial cells, even neurons and glial cells of the nervous system, also have the functions of immune regulation and promoting hematopoietic reconstruction, making it an ideal treatment for multi-system diseases "Practical Stem Cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08
Inventor 王福生施明徐若男
Owner THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
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