Human umbilical cord mesenchymal stem cell culture solution and culture method thereof

A technology of quality stem cells and culture fluid, applied in cell culture media, cell culture active agents, animal cells, etc., can solve problems such as adverse reactions, and achieve the effects of avoiding adverse reactions, avoiding moral disputes, and high purity

Inactive Publication Date: 2017-04-26
SHANGHAI AISAER BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Therefore, the present invention aims at the technical problem that the human umbilical cord mesenchymal stem cell culture system in the prior art contains human serum and/or animal-derive

Method used

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  • Human umbilical cord mesenchymal stem cell culture solution and culture method thereof
  • Human umbilical cord mesenchymal stem cell culture solution and culture method thereof
  • Human umbilical cord mesenchymal stem cell culture solution and culture method thereof

Examples

Experimental program
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Effect test

Embodiment 1～3

[0061] Examples 1-3 Culture of Human Umbilical Cord Mesenchymal Stem Cells Using Tissue Block Adhesion Method

[0062] training process

[0063] 1. Cord Tissue Processing

[0064] cut the umbilical cord into n 1 cm section, washed 3 times quickly with 75% alcohol, in Ca-free solution containing GA-1000 2+ and Mg 2+ Try to remove the three blood vessels in the DPBS; gently scrape the inner layer twice with a scraper to remove the single layer of endothelial cells, and then use Ca-free solution containing GA-1000 2+ and Mg 2+ Wash 3 times with DPBS, move the treated umbilical cord into a new petri dish, cut the umbilical cord into n 2 mm 3 umbilical cord fragments.

[0065] 2. Primary culture of human umbilical cord mesenchymal stem cells

[0066] Move the cut umbilical cord fragments into a new petri dish, and make them scatter and paste on the petri dish piece by piece, and the density of the entire petri dish reaches C 3 %. Open the lid of the culture dish and let i...

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Abstract

The invention discloses a human umbilical cord mesenchymal stem cell culture solution. The culture solution comprises a concentrated lysate, an Iscove's modified medium and an insulin transferrin selenite supplement liquid according to a volume ratio of (4-6):100:(0.8-1.2), and also comprises 4-6 ng/mL of a trypsin inhibitor, 12-18 ng/mL of a recombinant human epidermal growth factor and 12-18 ng/mL of a basic fibroblast growth factor. The culture solution is adopted to carry out adherent culture on umbilical cord tissues; a TrypLE digestion juice is adopted to carry out digestion; and subculturing is carried out. Human umbilical cord mesenchymal stem cells obtained in the invention have high purity, and can be directly used in clinic researches; and a culture method has a high amplification efficiency, is convenient for material drawing, and avoids moral dispute. A culture system of the human umbilical cord mesenchymal stem cells contains no human serum or animal-derived protein components, greatly reduces adverse effects caused by introduction of exogenous proteins in clinic treatment, and can be directly applied to the clinic researches.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a human umbilical cord mesenchymal stem cell culture medium and a culture method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells derived from the mesoderm and ectoderm in early development, and were originally found in the bone marrow. Under specific induction conditions in vitro, mesenchymal stem cells can differentiate into fat, cartilage, bone, muscle, tendon, nerve, liver, myocardium, pancreatic β cells, and endothelial cells. Potential for multidirectional differentiation. [0003] The main sources of umbilical cord mesenchymal stem cells are: [0004] 1) Bone marrow source: The number and proliferation and differentiation potential of adult bone marrow-derived mesenchymal stem cells decrease with age, and the virus infection rate is high, and the collection of donor mesenchymal stem cells requires ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N5/005C12N2500/25C12N2500/95C12N2501/11C12N2501/115C12N2501/734
Inventor 高歌杨帆
Owner SHANGHAI AISAER BIOTECH CO LTD
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