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Method for culturing mesenchymal stem cell from umbilical cord and application thereof

A technology of mesenchymal stem cells and umbilical cord, which is applied in the field of culture and application of umbilical cord-derived mesenchymal stem cells, and can solve the problems of increasing the risk of animal virus infection in animal components

Inactive Publication Date: 2015-06-24
JILIN TUO HUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research and clinical practice of regenerative medicine require a large number of seed cells. At present, most of the cell cultures are supplemented with fetal bovine serum, which increases the influence of animal components and the risk of various animal virus infections.

Method used

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  • Method for culturing mesenchymal stem cell from umbilical cord and application thereof
  • Method for culturing mesenchymal stem cell from umbilical cord and application thereof
  • Method for culturing mesenchymal stem cell from umbilical cord and application thereof

Examples

Experimental program
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preparation example Construction

[0031] Example 1 Preparation method of umbilical cord-derived mesenchymal stem cells

[0032] 1. Main reagents and consumables:

[0033] a-MEM medium (Hyclone Company), serum substitute (Daktronics Company, Helios UltraGROTM). Cell dissociation enzyme TrypLETMSelect (Life Company), normal saline (Juneng Pharmaceutical), T75 cell culture flask (NEST Company, polystyrene material), T182 cell culture flask (JET Company), for flow cytometry detection Antibodies CD14, CD19, CD34, CD45, CD73, CD90, CD105, HLA-DR and their isotype controls were purchased from ebioscience, USA. 10ml small beaker, self-made stainless steel stirring rod: made thin, a certain length, wherein, its diameter is 3-5mm, length 20-30cm, and can penetrate into the bottom corner of the culture bottle; one side of the stainless steel stirring rod is The cylindrical shape and the flat shape on the other side are convenient for pushing the muddy umbilical cord tissue and evenly distributing it into a dot matrix, ...

Embodiment 2

[0054] Example 2 The effect of using umbilical cord-derived mesenchymal stem cells in immune diseases

[0055] Zhao Moumou, female, 54 years old, was diagnosed with idiopathic thrombocytopenic purpura. After treatment with hormones, gamma balls, and splenectomy, the platelets fluctuated between (10-30)×10^9 / L. The normal range of platelets is (100-300)×10^9 / L. 2012-11-09 Infusion of umbilical cord-derived mesenchymal stem cells, one week later, platelets increased to 75×10^9 / L, and he was discharged from the hospital. One month later, the outpatient follow-up showed that the platelets were at a normal level of 128×10^9 / L. No other drugs were used during the period.

[0056] Treatment method: Digest 15 bottles of umbilical cord-derived mesenchymal stem cells (T182) cultured to the P4 generation with cell dissociating enzyme. After collection, wash twice with normal saline. Sent to department for intravenous infusion.

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Abstract

The invention relates to a method for culturing a mesenchymal stem cell from an umbilical cord and an application thereof. Particularly the mesenchymal stem cell is obtained by performing isolated culture on an umbilical cord tissue discarded by a full-term caesarean puerpera. A simple method for culturing plenty of seed cells is provided for treating a tissue injury disease and researching a regenerative medicine through the mesenchymal stem cell. The method is strong in practicability without animal content and is safe to use.

Description

technical field [0001] The present invention relates to a primary extraction method of umbilical cord-derived mesenchymal stem cells and a subculture operation method for obtaining a large number of mesenchymal stem cells not involving animal components, and the obtained mesenchymal stem cells in Use in autoimmune diseases. Background technique [0002] The self-renewal and multiple differentiation potentials of mesenchymal stem cells are more and more widely used in tissue damage and repair. In addition, a variety of cytokines secreted by it have an important regulatory effect on the endocrine and immune systems of the human body. The source of mesenchymal stem cells has developed from bone marrow, which was first discovered, to various tissues such as umbilical cord, umbilical cord blood, teeth, placenta, embryonic liver, and fat. However, among these sources, bone marrow and adipose-derived materials bring pain to patients, and the mesenchymal content of bone marrow is ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P37/08A61P7/04C12M1/24C12M1/02
Inventor 毕薇薇
Owner JILIN TUO HUA BIOTECH
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