Method for inducing differentiation of porcine dedifferentiated adipocytes to form skeletal muscle cells
A technology of skeletal muscle cells and adipocytes, applied in the field of using porcine dedifferentiated adipocytes to induce differentiation into skeletal muscle cells, can solve the problems of not having the biological characteristics and functions of porcine DFAT cells, not disclosing DFAT cells, and not providing experimental data, etc. Achieve the effect of simple and convenient induction procedure, convenient sampling and abundant sources
Inactive Publication Date: 2016-10-26
HUAZHONG AGRI UNIV
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[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 DFAT cells, and does not The obtained porcine DFAT cells were studied and verified for their biological characteristics and functions, and no relevant experimental data was provided to support the evidence that the isolated DFAT cells were preadipocytes, nor were they disclosed in the examples of the patent publication. Information on the biological identification of isolated DFAT cells, especially without any experimental support and results for the differentiation of porcine DFAT cells into myogenic direction
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Embodiment 1
[0053] 1. Isolation and purification of porcine mature adipocytes and culture by "ceiling" method
[0054] (1) Separation and purification of pig mature adipocytes: Take 5-day-old "Landrace" boars (or "Landrace boars" for short, provided by the Experimental Breeding Pig Farm of Huazhong Agricultural University, which is a conventional variety), and treat them with 75% alcohol. After the whole body of the pig was sterilized twice, it was sacrificed by exsanguination from the carotid artery, and the blood was drained as much as possible. Subsequently, the whole body of the piglet was wiped with alcohol cotton containing 75% alcohol for disinfection. Move the piglet into the ultra-clean workbench, use sterilized scissors and tweezers under aseptic conditions, take the subcutaneous fat of the back of the piglet and place it in a place containing 1% double antibody (anti-penicillin-streptomycin, purchased from Hangzhou Jinuo Biomedical Technology Co., Ltd. ) phosphate-buffered sal...
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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 al 2006). [0003] However, due to the small number of satellite cells isolated from muscle tissue in ...
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IPC IPC(8): C12N5/077
Inventor 彭健彭学武
Owner HUAZHONG AGRI UNIV
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