Preparation and storage of mesenchymal stem cells for clinical treatment

A technology of mesenchymal stem cells and umbilical cords, applied in the field of preparation and storage of human umbilical cord mesenchymal stem cells, can solve the problems of increasing pollution of hematopoietic stem cells, increasing the difficulty of umbilical cord processing, etc., and achieve good application prospects, excellent results, and low cost effects

Inactive Publication Date: 2011-08-17
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

[0005] 1 After the umbilical cord is collected, the umbilical cord and arterial blood coagulate, which increases the difficulty of umbilical cord processing, and at the same time, too many blood cells increase the chance of hematopoietic stem cell contamination

Method used

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  • Preparation and storage of mesenchymal stem cells for clinical treatment
  • Preparation and storage of mesenchymal stem cells for clinical treatment
  • Preparation and storage of mesenchymal stem cells for clinical treatment

Examples

Experimental program
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Embodiment 1

[0027] 1. Under aseptic conditions, the umbilical cord of a normal child or caesarean section was collected, about 30 cm long, and the umbilical cord tissue was preserved with normal saline containing 0.2% gentamicin and 0.54% anticoagulant. Carry out pathogenic microbiology and virology testing at the same time, after being fully qualified, proceed to the following steps.

[0028] 2. Place the umbilical cord in physiological saline containing 0.2% gentamicin, wash it 2-3 times repeatedly, cut off the umbilical vein and arteries with sterile scissors, and remove the visible vascular tissue and possible blood clots. Use scissors to cut the umbilical cord to a size of 2-3cm, collect the tissue pieces and place them in a 50ml centrifuge tube, centrifuge at a high speed at 2000 rpm for 10 minutes, discard the supernatant, and add 0.2% gentamicin Sufficient normal saline, fully shake the precipitated tissue pieces to loosen them, and centrifuge at a high speed at 2000 rpm for 10 mi...

Embodiment 2

[0037]1. Under sterile conditions, collect the umbilical cord of a normal or cesarean section fetus, about 40 cm long, and preserve the umbilical cord tissue with normal saline containing 0.2% gentamicin and 0.54% anticoagulant. Carry out pathogenic microbiology and virology testing at the same time, after being fully qualified, proceed to the following steps.

[0038] 2. Place the umbilical cord in physiological saline containing 0.2% gentamicin, wash it 2-3 times repeatedly, cut off the umbilical vein and arteries with sterile scissors, and remove the visible vascular tissue and possible blood clots. Use scissors to cut the umbilical cord to a size of 2-3cm, collect the tissue pieces and place them in a 50ml centrifuge tube, centrifuge at a high speed at 2000 rpm for 10 minutes, discard the supernatant, and add 0.2% gentamicin Sufficient normal saline, fully shake the precipitated tissue pieces to loosen them, and centrifuge at a high speed at 2000 rpm for 10 minutes.

[00...

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Abstract

The invention relates to preparation and storage of mesenchymal stem cells for clinical treatment. The preparation method comprises the following steps: storing a physiological saline umbilical cord containing 0.2% of genramicin and 0.54% of anticoagulant; shearing the umbilical cord to 2-3cm; centrifuging, suspending, and centrifuging; directly adhering and carrying out explant-culture; adding an alpha-minimum essential medium (MEM) containing 15% of fetal calf serum and 0.2% of genramicin, and culturing; after cell fusion, digesting the cells with digestive juice containing 0.05% of pancreatin/0.2mM ethylene diamine tetraacetic acid (EDTA); carrying out passage in a ratio of 1:3 to 4-5 generations, so as to obtain a large amount of mesenchymal stem cells; and putting a frozen storing liquid (the ratio of medium to fetal calf serum to dimethylsulfoxide is 5:4:1) in the mesenchymal stem cells, placing in a refrigerator of minus 70 DEG C overnight and then storing in liquid nitrogen, wherein the freezing temperature of the liquid nitrogen is minus 196 DEG C.

Description

technical field [0001] The invention relates to the preparation of a medical product, in particular to the preparation and preservation 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 王福生施明徐若男
Owner THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
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