Serum-free polypeptide composition for promoting proliferation of mesenchymal stem cells

A technology of polypeptide composition and stem cells, applied in the field of cell proliferation, can solve the problems of inability to meet the requirements of mass production, high price, high risk of introduction of animal-derived proteins, etc. Effects of chemical properties and immunophenotypic stability

Active Publication Date: 2019-06-28
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These media are not only expensive and cannot meet the requirements of mass production, but also need to coat th

Method used

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  • Serum-free polypeptide composition for promoting proliferation of mesenchymal stem cells
  • Serum-free polypeptide composition for promoting proliferation of mesenchymal stem cells
  • Serum-free polypeptide composition for promoting proliferation of mesenchymal stem cells

Examples

Experimental program
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preparation example Construction

[0063] The preparation of the polypeptide composition described in the present application can be prepared according to the methods well known to those skilled in the art. After the specific time is mixed, the components are filtered and sterilized in the filter membrane, and then added to the α-MEM basal medium and mixed evenly, that is, Serum-free medium for promoting proliferation of mesenchymal stem cells (UC-MSCs) was obtained.

[0064] The mesenchymal stem cell-promoting polypeptide composition provided by the present invention has clear chemical composition, no animal source, no serum, can realize the rapid proliferation of UC-MSCs, solve the problem of insufficient number of cells, and maintain the biological characteristics and immunity of mesenchymal stem cells Phenotypic stability.

Embodiment 1

[0067] Example 1 Preparation of serum-free medium

[0068] 1) Based on α-MEM basal medium, prepare serum-free medium according to the following raw material ratio:

[0069] Non-essential amino acids: 1% by volume;

[0070] Glutamine: 1~4 mM;

[0071] Lipid mixture: 1vol%;

[0072] ITS: 1vol%;

[0073] Recombinant human serum albumin: 2.5g / L;

[0074] Recombinant human epidermal growth factor: 20µg / L;

[0075] Recombinant human fibronectin: 10µg / L;

[0076] L-glutathione: 2mg / L;

[0077] β-mercaptoethanol: 2mg / L;

[0078] Tripeptide-1: 10µg / L;

[0079] Tripeptide-2: 10µg / L;

[0080] Hexapeptide-9: 10µg / L;

[0081] Palmitoyl Hexapeptide-12: 10µg / L;

[0082] Laminin-derived peptide: 50µg / L;

[0083] α-MEM basal medium 10.2g / L;

[0084] The above-mentioned components were dissolved in water at room temperature and fully dissolved to obtain the polypeptide composition of the present application as the experimental group 2; and then sterilized by filtration through a 0....

Embodiment 2

[0144] Example 2 Morphological observation and activity detection of UC-MSCs

[0145] Select the P3 generation UC-MSCs to carry out the experiment, and the UC-MSCs are divided into 1×10 4 / cm 2 The density was inoculated in T25 culture flasks, with 3 replicates in each group; placed in 5% CO 2 Culture in an incubator at 37°C; collect images of UC-MSCs after 48 hours of culture, and the results are as follows: figure 1 as shown, figure 1 Middle panel A is the morphological diagram of UC-MSCs in control group 1, panel B is the morphological diagram of UC-MSCs in control group 2, panel C is the morphological diagram of UC-MSCs in experimental group 1, and panel D is the morphological diagram of UC-MSCs in experimental group 2 Morphological diagram, Figure E is the morphological diagram of UC-MSCs of experimental group 3.

[0146] Depend on figure 1 It can be seen that the UC-MSCs in each group grew in a single layer of adherent, and most of the cells were long spindle-shaped...

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Abstract

The invention provides a serum-free polypeptide composition for promoting proliferation of mesenchymal stem cells. Each liter of the serum-free polypeptide composition mainly comprises 10-100 micrograms of tripeptide-1, 1-20 micrograms of tripeptide-2, 1-20 micrograms of hexapeptide-9, 1-20 micrograms of palmitoyl hexapeptide-12 and 10-100 micrograms of laminin-derived peptide. The serum-free polypeptide composition contains clear chemical components without animal sources or serum, and can realize rapid proliferation of mesenchymal stem cells, solve the problem of insufficient cell quantity and maintain the biological characteristics and immunophenotypic stability of mesenchymal stem cells.

Description

technical field [0001] The invention relates to the technical field of cell proliferation, in particular to a serum-free polypeptide composition for promoting proliferation of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (Mesenchymal stem cells, MSCs) are derived from the mesoderm in the early stage of development. tissue, umbilical cord, cord blood and placenta. MSCs have high self-renewal ability and multi-directional differentiation potential, can be cultured and expanded in vitro, not only can support the growth of hematopoietic stem cells, but also have the function of immune regulation; under different induction conditions, they can differentiate into bone, cartilage, muscle in vitro , nerve, myocardium, endothelium and fat, etc., still have multi-directional differentiation potential after continuous subculture and cryopreservation, and can be used as ideal seed cells for the repair of tissue and organ damage caused by aging and diseas...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N2500/90C12N2501/998C12N5/0668C12N2500/36C12N2533/52C12N2501/11C12N5/0662C12N2500/25C12N2500/30C12N2500/32C12N2500/44
Inventor 陈海佳葛啸虎王小燕戚康艺姜交华
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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