Serum-free culture medium for human umbilical cord MSCs (mesenchymal stem cells)

A technology of serum-free medium and mesenchymal stem cells, which is applied in the field of cell engineering, can solve the problems of insufficient number of cells, achieve the effect of solving insufficient number of cells, solving biological safety problems, and maintaining characteristics

Inactive Publication Date: 2017-08-15
王晓柯
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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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Preparation of serum-free medium for human umbilical cord mesenchymal stem cells

[0064] The concentration of each raw material is as follows, each concentration unit is mg / L unless otherwise specified:

[0065] L-Arginine 200;

[0066] CuSO 4 ·5H 2 O 0.002;

[0067] L-Asparagine 50;

[0068] L-Aspartic Acid 20;

[0069] L-glutamic acid 20;

[0070] Ni(NO 3 ) 2 ·6H 2 O 0.0001;

[0071] L-glutamine 300;

[0072] ZnSO 4 ·7H 2 O 0.3;

[0073] Glycine 10;

[0074] CoCl 2 ·6H 2 O 0.005;

[0075] L-histidine 20;

[0076] NaSiO 3 ·9H 2 O 0.005;

[0077] L-Isoleucine 50;

[0078] Na 3 VO 4 ·12H 2 O 0.002;

[0079] L-lysine hydrochloride 40;

[0080] SnCl 2 ·2H 2 O 0.00005;

[0081] L-methionine 20;

[0082] Na 2 SeO 3 0.005;

[0083] L-phenylalanine 20;

[0084] FeSO 4 ·7H 2 O 1.0;

[0085] L-Proline 20;

[0086] Glucose 3000;

[0087] L-serine 30;

[0088] Vitamin C 0.500;

[0089] L-threonine 20;

[0090] P-Hydroxybenzoic acid 1.0;

[0091] L-tryptophan 10;

[0092] Sodium pyruvate 300;

[0093] L-...

Embodiment 2

[0115] Example 2 Preparation of serum-free medium for human umbilical cord mesenchymal stem cells

[0116] The concentration of each raw material is as follows, each concentration unit is mg / L unless otherwise specified:

[0117] L-Arginine 100;

[0118] CuSO 4 ·5H 2 O 0.0005;

[0119] L-Asparagine 75;

[0120] L-Aspartic acid 30;

[0121] L-glutamic acid 10;

[0122] Ni(NO 3 ) 2 ·6H 2 O 0.00002;

[0123] L-glutamine 500;

[0124] ZnSO 4 ·7H 2 O 0.6;

[0125] Glycine 5;

[0126] CoCl 2 ·6H 2 O 0.001;

[0127] L-histidine 30;

[0128] NaSiO 3 ·9H 2 O 0.01;

[0129] L-Isoleucine 25;

[0130] Na 3 VO 4 ·12H 2 O 0.0005;

[0131] L-lysine hydrochloride 60;

[0132] SnCl 2 ·2H 2 O 0.0001;

[0133] L-methionine 10;

[0134] Na 2 SeO 3 0.002;

[0135] L-phenylalanine 30;

[0136] FeSO 4 ·7H 2 O 1.6;

[0137] L-Proline 10;

[0138] Glucose 1000;

[0139] L-serine 45;

[0140] Vitamin C 0.704;

[0141] L-threonine 10;

[0142] P-Hydroxybenzoic acid 0.5;

[0143] L-tryptophan 15;

[0144] Sodium pyruvate 550;

[0145] L-...

Embodiment 3

[0167] Example 3 Preparation of serum-free medium for human umbilical cord mesenchymal stem cells

[0168] The concentration of each raw material is as follows, each concentration unit is mg / L unless otherwise specified:

[0169] L-Arginine 300;

[0170] CuSO 4 ·5H 2 O 0.005;

[0171] L-Asparagine 25;

[0172] L-Aspartic Acid 10;

[0173] L-glutamic acid 30;

[0174] Ni(NO 3 ) 2 ·6H 2 O 0.0002;

[0175] L-glutamine 0;

[0176] ZnSO 4 ·7H 2 O 0.06;

[0177] Glycine 15;

[0178] CoCl 2 ·6H 2 O 0.008;

[0179] L-histidine 10;

[0180] NaSiO 3 ·9H 2 O 0.001;

[0181] L-Isoleucine 75;

[0182] Na 3 VO 4 ·12H 2 O 0.005;

[0183] L-lysine hydrochloride 20;

[0184] SnCl 2 ·2H 2 O 0.00001;

[0185] L-methionine 30;

[0186] Na 2 SeO 3 0.01;

[0187] L-phenylalanine 10;

[0188] FeSO 4 ·7H 2 O 0.2;

[0189] L-Proline 30;

[0190] Glucose 4000;

[0191] L-serine 15;

[0192] Vitamin C 0.176;

[0193] L-threonine 30;

[0194] P-Hydroxybenzoic acid 1.5;

[0195] L-tryptophan 5;

[0196] Sodium pyruvate 55;

[0197] L-vali...

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PUM

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Abstract

The invention discloses a serum-free culture medium for human umbilical cord MSCs (mesenchymal stem cells). The serum-free culture medium comprises the major ingredients of amino acid, inorganic salt ingredients, growth factors and albumin ingredients. The serum-free culture medium for human umbilical cord MSCs does not contain any serum ingredients; in addition, the ingredients are clear; the foreign protein pollution and pathogenic microorganism risks are avoided; the biosecurity problem of the MSCs clinic application is solved. When the serum-free culture medium provided by the invention is used for culturing the human umbilical cord MSCs, the high-quantity high-purity safe and high-quality umbilical cord MSCs can be obtained in vitro; meanwhile, the cell survival rate can be improved; the stem cell characteristics are maintained. The problem of insufficient cell quantity can be solved; the cell number, the survival rate and the immunophenotyping in each batch are more stable; the relevant index requirements are met.

Description

Technical field [0001] The invention relates to a serum-free medium for human umbilical cord mesenchymal stem cells, which belongs to the technical field of cell engineering. Background technique [0002] Mesenchymal stem cells (MSCs) are derived from the mesenchymal stem cells and are also called multipotent mesenchymal stromal cells. They are a type of pluripotent stem cells. They are first found in the bone marrow and have three types. The multi-directional differentiation potential of germ layer cells has low immunogenicity, does not stimulate allogeneic immune responses, is not killed by cytotoxic T cells and NK cells, and has immunoregulatory effects. MSCs can differentiate into different types of cells such as bone, fat, cartilage, muscle and liver cells under the corresponding induction conditions. Umbilical cord mesenchymal stem cells are a type of cell population that exists in the human umbilical cord and has multidirectional differentiation potential and self-renewal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2500/10C12N2500/12C12N2500/20C12N2500/22C12N2500/25C12N2500/30C12N2500/32C12N2500/34C12N2500/36C12N2500/38C12N2500/44C12N2500/46C12N2500/90C12N2501/11C12N2501/15C12N2501/39C12N2501/71C12N2501/998
Inventor 王晓柯
Owner 王晓柯
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