Method of forming skeletal muscle cell by induction differentiation by utilization of pig dedifferentiated adipocyte

A technology for skeletal muscle cells and adipocytes is applied in the field of using pig dedifferentiated adipocytes to induce differentiation into skeletal muscle cells, which can solve the problems of lack of biological characteristics and functions of pig DFAT cells, no disclosure of DFAT cells, and no experimental data provided. To achieve the effect of simple and convenient induction procedure, convenient sampling and rich sources

Inactive Publication Date: 2014-10-29
HUAZHONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0006] The patent application literature related to the present invention is the method reported by the application number 201110002340.4 (name of invention: a culture method for rapid dedifferentiation of porcine mature adipocytes into preadipocytes). Mature adipocytes were isolated from tissues, and porcine DFAT cells were obtained through the "ceiling" culture method. This document discloses the method and path of using DFAT cells as preadipocytes, but only discloses the separation method for obtaining porcine

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  • Method of forming skeletal muscle cell by induction differentiation by utilization of pig dedifferentiated adipocyte
  • Method of forming skeletal muscle cell by induction differentiation by utilization of pig dedifferentiated adipocyte
  • Method of forming skeletal muscle cell by induction differentiation by utilization of pig dedifferentiated adipocyte

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[0052] Example 1

[0053] 1. Separation and purification of mature pig fat cells and "ceiling" method culture

[0054] (1) Isolation and purification of mature pig fat cells: Take 5-day-old "Landrace pig" boar (or simply "Landrace boar", provided by the experimental pig farm of Huazhong Agricultural University, which is a conventional breed), and use 75% alcohol to After the whole body was disinfected twice, the pigs were put to death by bleeding from the carotid artery, and the blood was drained as much as possible. Afterwards, wipe the piglets with an alcohol cotton containing 75% alcohol for disinfection. Move the piglet to the ultra-clean workbench, use sterilized scissors and tweezers under aseptic conditions, take the subcutaneous fat on the back of the piglet and place it with 1% double antibody (anti-penicillin-streptomycin, purchased from Hangzhou Jinuo Biomedical Technology Co., Ltd. ) In a phosphate buffered saline (PBS) dish, rinse with PBS for 4-5 times to ensure ste...

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Abstract

The invention belongs to the technical field of animal cell culturing, and particularly relates to a method of forming a skeletal muscle cell by induction differentiation by utilization of a pig dedifferentiated adipocyte. A mature adipocyte of pig subcutaneous adipose tissues is separated by a collagenase digestion method, and is dedifferentiated to form the dedifferentiated adipocyte (DFAT cell). The DFAT cell has functions similar to that of mesenchymal stem cells (MSCs) and can be efficiently multiplicated in vitro and can introduce multi-directional differentiation. In addition, by utilization of induction of the galectin-1 protein in vitro, the pig DFAT cell is dedifferentiated to form a multinucleate cell capable of expressing a skeletal muscle cell specific marker molecule.

Description

technical field [0001] The invention belongs to the technical field of in vitro culture of animal cells, and in particular relates to a method for inducing differentiation of porcine dedifferentiated adipocytes (DFAT) to form skeletal muscle cells. Background technique [0002] Pig muscle tissue is an important production trait in pig production. The skeletal muscle cells that form muscle tissue are multinucleated cells formed from mesoderm stem cells after finalization and differentiation. Unfortunately, no porcine-derived myoblast or stem cell lines have been established for the study of stem cell commitment and differentiation into skeletal muscle cells. Mesenchymal stem cells (mesenchymal stem cells, referred to as MSCs, the same below) and satellite cells derived from adult tissues are commonly used models for myogenic differentiation of primary stem cells (Dezawa et al2006). [0003] However, due to the small number of satellite cells isolated from muscle tissue in v...

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Application Information

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IPC IPC(8): C12N5/077
Inventor 彭健彭学武
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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