Serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells

A serum-free medium and large-scale culture technology, which is applied in the field of cell engineering, can solve the problems of insufficient number of cells, etc., and achieve the effect of solving insufficient number of cells, clear components, and stable immune phenotype

Inactive Publication Date: 2019-02-22
YOCON BIOLOGY TECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention can solve the problem of insufficient cell quantity, and the cell quantity, survival rate and immune phenotype of each batch are more stable

Method used

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  • Serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells
  • Serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Preparation of serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells

[0067] The concentration of each raw material is as follows, and the unit of each concentration is mg / L unless otherwise specified:

[0068] L-Arginine 200;

[0069] CuSO 4 ·5H 2 O 0.002;

[0070] L-Asparagine 50;

[0071] L-aspartic acid 20;

[0072] L-glutamic acid 20;

[0073] Ni (NO 3 ) 2 ·6H 2 O 0.0001;

[0074] L-Glutamine 300;

[0075] ZnSO 4 ·7H 2 O 0.3;

[0076] Glycine 10;

[0077] CoCl 2 ·6H 2 O 0.005;

[0078] L-histidine 20;

[0079] NaSiO 3 9H 2 O 0.005;

[0080] L-Isoleucine 50;

[0081] Na 3 VO 4 12H 2 O 0.002;

[0082] L-lysine hydrochloride 40;

[0083] SnCl 2 2H 2 O 0.00005;

[0084] L-methionine 20;

[0085] Na 2 SeO 3 0.005;

[0086] L-phenylalanine 20;

[0087] FeSO 4 ·7H 2 O 1.0;

[0088] L-proline 20;

[0089] Glucose 3000;

[0090] L-serine 30;

[0091] Vitamin C 0.500;

[0092] L-threonin...

Embodiment 2

[0117] Example 2 Preparation of serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells

[0118] The concentration of each raw material is as follows, and the unit of each concentration is mg / L unless otherwise specified:

[0119] L-Arginine 100;

[0120] CuSO 4 ·5H 2 O 0.0005;

[0121] L-asparagine 75;

[0122] L-aspartic acid 30;

[0123] L-glutamic acid 10;

[0124] Ni (NO 3 ) 2 ·6H 2 O 0.00002;

[0125] L-Glutamine 500;

[0126] ZnSO 4 ·7H 2 O 0.6;

[0127] Glycine 5;

[0128] CoCl 2 ·6H 2 O 0.001;

[0129] L-histidine 30;

[0130] NaSiO 3 9H 2 O 0.01;

[0131] L-isoleucine 25;

[0132] Na 3 VO 4 12H 2 O 0.0005;

[0133] L-lysine hydrochloride 60;

[0134] SnCl 2 2H 2 O 0.0001;

[0135] L-Methionine 10;

[0136] Na 2 SeO 3 0.002;

[0137] L-Phenylalanine 30;

[0138] FeSO 4 ·7H 2 O 1.6;

[0139] L-proline 10;

[0140] Glucose 1000;

[0141] L-serine 45;

[0142] Vitamin C 0.704;

[0143] L-threonin...

Embodiment 3

[0168] Example 3 Preparation of serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells

[0169] The concentration of each raw material is as follows, and the unit of each concentration is mg / L unless otherwise specified:

[0170] L-Arginine 300;

[0171] CuSO 4 ·5H 2 O 0.005;

[0172] L-asparagine 25;

[0173] L-Aspartate 10;

[0174] L-glutamic acid 30;

[0175] Ni (NO 3 ) 2 ·6H 2 O 0.0002;

[0176] L-glutamine 0;

[0177] ZnSO 4 ·7H 2 O 0.06;

[0178] Glycine 15;

[0179] CoCl 2 ·6H 2 O 0.008;

[0180] L-histidine 10;

[0181] NaSiO 3 9H 2 O 0.001;

[0182] L-isoleucine 75;

[0183] Na 3 VO 4 12H 2 O 0.005;

[0184] L-lysine hydrochloride 20;

[0185] SnCl 2 2H 2 O 0.00001;

[0186] L-methionine 30;

[0187] Na 2 SeO 3 0.01;

[0188] L-Phenylalanine 10;

[0189] FeSO 4 ·7H 2 O 0.2;

[0190] L-proline 30;

[0191] Glucose 4000;

[0192] L-serine 15;

[0193] Vitamin C 0.176;

[0194] L-threonine 30; ...

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Abstract

The invention discloses a serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells. The medium mainly includes amino acid, an inorganic salt component, a growth factorand an albumin component. The serum-free medium does not contain any serum component, has clear components, overcomes the danger of heterologous protein contamination and pathogenic microorganisms, and solves the biosafety problem of MSCs clinical application. The human umbilical cord mesenchymal stem cells can be cultured invitro in large scale by using the serum-free medium. The umbilical cord mesenchymal stem cells with high volume, high purity, safety and high quality can be obtained. At the same time, the cell survival rate can be improved, and the characteristics of stem cells can be maintained. The serum-free medium can solve the problem of insufficient cell number, the cell number, the survival rate and the immunophenotype of each batch are more stable, and the requirements for relevant indexes can be met.

Description

technical field [0001] The invention relates to a serum-free medium for large-scale culture of human umbilical cord mesenchymal stem cells, belonging to the technical field of cell engineering. Background technique [0002] Mesenchymal stem cells (MSCs) are derived from mesoderm, also known as multipotent mesenchymal stromal cells (multipotent mesenchymal stromal cells). The multilineage differentiation potential of germ layer cell differentiation has low immunogenicity, does not stimulate allogeneic immune response, is not killed by cytotoxic T cells and NK cells, and has immunomodulatory effects. Under the corresponding induction conditions, MSCs can differentiate into different types of cells such as bone, fat, cartilage, muscle and liver cells. Umbilical cord mesenchymal stem cells are a kind of cell population with multilineage differentiation potential and self-renewal ability existing in human umbilical cord. It is convenient to obtain materials and has a wide range...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2501/15C12N2511/00C12N2501/998C12N2501/71C12N2501/39C12N2500/22C12N2500/20C12N2500/25C12N2500/30C12N2500/12C12N2500/10C12N2500/44C12N2500/38C12N2500/46C12N2500/32C12N2500/36C12N2500/34C12N2501/115C12N2500/90
Inventor 张昕
Owner YOCON BIOLOGY TECH CO
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