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In-vitro culturing method for porcine skeletal muscle satellite cell (SMSC)

A satellite cell, in vitro culture technology, used in bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve problems such as low proliferation ability, hindering research on muscle tissue regeneration and development mechanism, and inability to meet gene transfer.

Inactive Publication Date: 2011-08-03
NORTHWEST A & F UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Pig SMSC isolation and culture methods mostly use trypsin, collagenase step-by-step or pronase protease one-step digestion. The SMSC isolated and cultured by these methods are mainly myoblasts and non-myoblasts (fibroblasts) with low proliferative ability, and the purity is low. Failure to meet the SMSC-mediated gene transfer hinders the research on the mechanism of muscle tissue regeneration and development, and also fails to meet the needs of cell transplantation for the treatment of muscle-related diseases

Method used

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  • In-vitro culturing method for porcine skeletal muscle satellite cell (SMSC)
  • In-vitro culturing method for porcine skeletal muscle satellite cell (SMSC)
  • In-vitro culturing method for porcine skeletal muscle satellite cell (SMSC)

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Embodiment Construction

[0031] The present invention will be described in detail below in conjunction with specific embodiments.

[0032] 1. Experimental animals

[0033] Healthy large white pigs aged 1-3 days were provided by the livestock and poultry ecological farm of Northwest Agriculture and Forestry University.

[0034] 2. Sample collection

[0035] The large white pigs were washed with 0.5% bromogeramine for 0.5 h, and then killed by electric shock. Then, on a sterile operating table, operate on the tendons at both ends, carefully and completely strip the Extensor Digitorum Longus (EDL), and at the same time, according to its special composition mechanism, further strip it into four small pieces from the tendon, In order to facilitate the full digestion of enzymes.

[0036] 3. Pretreatment of Petri dishes

[0037] Coat the culture plate with Matrigel and place it at 37 °C, 5% CO 2 more than 4 hours in the incubator. Before use, after irradiating with UV light for 30 min, wash with PBS fo...

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Abstract

The invention discloses an in-vitro culturing method for a porcine skeletal muscle satellite cell (SMSC). The method comprises the following steps of: collecting a sample; pretreating a culture dish; transplanting skeletal muscle fibers; removing single muscle fibers; subculturing the SMSC; identifying the SMSC; and detecting the purity of the SMSC. A simple and practical high-purity porcine SMSC in-vitro culturing method is established, so that materials are provided for the researches of gene transfer of SMSC mediation, muscle tissue regeneration and developmental mechanism, and a basis is laid for the treatment of muscle-related diseases through skeletal muscle single fibers or SMSC transplanting.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for culturing porcine skeletal muscle satellite cells (Skeletal Muscle Satellite Cell, SMSC) in vitro. Background technique [0002] The in vitro culture of skeletal muscle satellite cells (Skeletal Muscle Satellite Cell, SMSC) is currently mainly carried out on mice, rats, rabbits, etc. Pigs are highly similar to humans in anatomical structure, organ size and life cycle, and are the closest model animal to humans, and their research is increasingly favored by scholars. Pig SMSC isolation and culture methods mostly use trypsin, collagenase step-by-step or pronase protease one-step digestion. The SMSC isolated and cultured by these methods are mainly myoblasts and non-myoblasts (fibroblasts) with low proliferative ability, and the purity is low. The lack of SMSC-mediated gene transfer hinders the research on the mechanism of muscle tissue regeneration and development, and al...

Claims

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Application Information

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IPC IPC(8): C12N5/077
Inventor 杨公社刘月光史新娥沈清武杨秋梅高晓娟陈宗正
Owner NORTHWEST A & F UNIV
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