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Method for alleviating or improving vascular diseases by using cell therapeutic agent

A cell therapy and cell technology, applied in cardiovascular diseases, animal cells, vertebrate cells, etc., can solve problems such as difficulty in being suitable for clinical use, difficult to maintain, and decreased cell viability.

Active Publication Date: 2020-02-28
BOYALIFE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] The 2016110569836 (Y16069) case of the inventor's team discloses a method for preparing CD34-positive cells from umbilical cord mesenchymal stem cells. However, the composition containing CD34-positive cells described therein is not suitable for long-term storage and transportation. It was found that the cell viability of this composition would decrease significantly after being frozen and revived, so it was difficult to be suitable for clinical use, such as storage and transportation before clinical use (for example, keeping the cell viability for 3 months at -80°C) process) is difficult to maintain

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0179] Embodiment 1, the pretreatment of human umbilical cord sample

[0180] Under sterile conditions, take the near-fetal umbilical cord of normal full-term production, disinfect the surface of the umbilical cord tissue with alcohol, and cut the umbilical cord from the middle;

[0181] Wash thoroughly with PBS containing 100U / mL penicillin and 100U / mL streptomycin to remove residual blood stains;

[0182] Divide the umbilical cord into 0.5cm umbilical cord segments, carefully remove the arteries and veins, take 50mL centrifuge tubes to hold the umbilical cord segments, 1 segment / centrifuge tube;

[0183] In a centrifuge tube, cut the umbilical cord segment into pieces with a size of about 1mm×1mm×1mm, and add LG-DMEM culture solution dropwise during the cutting process to keep the tissue moist;

[0184] Wash twice with PBS buffer solution containing 100 U / mL penicillin and 100 U / mL streptomycin (pH7.2, sodium phosphate salt, phosphate root concentration 0.025M, if not spec...

Embodiment 2

[0185] Embodiment 2, enzymolysis

[0186] The umbilical cord fragments obtained in Example 1 were mixed with enzyme solution (0.1% type I collagenase, 0.1% trypsin, 0.1% hyaluronidase, 0.1% DNase, 0.02% EDTA; % represents the percentage of mass concentration), Digest at 37°C for 2h;

[0187] After digestion, centrifuge for 15 minutes with a centrifugal force of 400g and a temperature of 4±2°C;

[0188] Discard the mixed enzyme solution to obtain single cells derived from umbilical cord.

Embodiment 3

[0189] Embodiment 3, primary culture of cells

[0190] The cells obtained from the treatment in Example 2 were divided into 1×10 6 The density of cells / ml was inoculated into T-25 culture flasks, placed in 20ml of RPMI1640 culture medium supplemented with 10% (v / v) fetal bovine serum, placed at 37°C, 5% CO 2 (v / v), incubating and cultivating for 3 days in an incubator with 95% humidity;

[0191] Change the medium for the first time after 3 days to remove unattached cells;

[0192] The culture medium was replaced every 48 h thereafter.

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PUM

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Abstract

The invention relates to a method for alleviating or improving vascular diseases by using a cell therapeutic agent of CD34 positive cells. On the one hand, the related cell therapy composition comprises CD34+ cells and a pharmaceutically-acceptable carrier, for example, the composition comprises CD34+ cells, sodium chloride and water for injection, wherein the density of the CD34+ cells is 1*106 cells / ml to 5*106 cells / ml, and the mass volume percentage of the sodium chloride is 0.8% to 1.0% or 0.9%. The invention further relates to a preparation method of the cell therapeutic agent containingCD34 positive cells and an application of the cell therapeutic composition in preparing drugs for relieving or improving vascular diseases, wherein the vascular diseases are peripheral arterial diseases caused by limb arterial stenosis and occlusion or diabetes, preferably lower limb arterial diseases caused by limb arterial stenosis and occlusion or diabetes. The method shows an excellent technical effect.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to the field of using cell biology technology to treat clinical diseases. Specifically, the present invention relates to a method for preparing CD34 from umbilical cord-derived mesenchymal stem cells + The cell method also relates to a cell therapy composition prepared from such cells. The cell therapy composition of the present invention can be used to alleviate or improve vascular disease, such as limb arterial stenosis and occlusion or peripheral arterial disease caused by diabetes; especially limb Lower extremity arterial disease caused by arterial stenosis and occlusion or diabetes. Background technique [0002] Stem cells mediate regeneration and the transmission of genetic information to subsequent cell generations. They can self-renew and produce differentiated progeny. In recent years, advances have been made in our understanding of the molecular mechanisms underlying the inte...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61P9/14C12N5/078
CPCA61K35/28A61P9/14C12N5/0634
Inventor 肖海蓉刘冰汤乐
Owner BOYALIFE
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