Dissertation > Excellent graduate degree dissertation topics show

Isolation and Culture of Porcine Skeletal Muscle Satellite Cells by Single Muscle Fibers and Their Myogenic Differentiation

Author: LiuYueGuang
Tutor: YangGongShe
School: Northwest University of Science and Technology
Course: Animal Genetic Breeding and Reproduction
Keywords: Skeletal muscle fibers Satellite cells Pax7 Pig
CLC: S828
Type: Master's thesis
Year: 2011
Downloads: 78
Quote: 0
Read: Download Dissertation

Abstract

Skeletal muscle is composed by terminally differentiated multinucleated muscle fibers. Therefore, the regeneration of skeletal muscle after birth depends on the library undifferentiated mononuclear muscle progenitor cells, satellite cells are the main component of the library. Satellite cells located between the surface and the basement membrane of the muscle fiber membrane, muscle-derived stem cells were in a resting state, has the potential of proliferation and differentiation of myoblasts, the stability of the environment skeletal muscle fiber hypertrophy and damage repair plays important role. Morphology, fusiform or ovoid, with a high nuclear cytoplasmic ratio and organelle that contains the characteristics of objects less. However, who is going to regulate satellite cell self-renewal or differentiation of understanding only just begun, currently considered resting satellite cells expressing Pax7 transcription factor, when it is activated, simultaneously express Pax7 and MyoD. When the proliferation complete, the majority of satellite cells down Pax7 expression, simultaneous differentiation. The other offspring satellite cells remained Pax7 expression but not the expression of MyoD, exit the cell cycle back to the resting state. Single muscle fiber isolation, culture has been widely used in research satellite cell gene expression, into the muscle development and muscle regeneration process. Isolated, cultured single muscle fiber to eliminate the impact of non-muscle cells, and to maintain its resting state, so it is very powerful, and domestic has not yet been reported. Therefore, this study by collagenase Ⅰ digestion from pig skeletal muscle separation complete a single muscle fibers and cultured on cell immune fluorescence identification of muscle fiber satellite cells, followed by the single muscle fiber free out of the satellite cells carry out cell immunofluorescence staining, mass culture, the growth curve drawing, induced differentiation of myoblasts and Western blot analysis of skeletal muscle satellite cells into muscle-specific protein expression. The main results are as follows: 1. Separated and cultured single muscle fibers adhered oval cells, and over time, the cells slow outward migration and proliferation of satellite cell-specific marker gene Pax7 and MyoD immunofluorescence staining of is positive, and the positive rate of 90% or more. Proliferation medium, add 1% chick embryo extract can significantly promote the proliferation of skeletal muscle satellite cells; at different periods, WB analysis of pig skeletal muscle satellite cell-specific protein found MyoD protein is highly expressed in the proliferative phase. Myogenin and MyHC expressed only to differentiation. Induced differentiation of myoblasts with the traditional DM Ⅰ differentiation medium cultured compare DM II differentiation medium more conducive to the confluence, differentiation to form multinucleated myotubes, and into the muscle-specific marker gene Myogenin and MyHC expression level was significantly upregulated. To sum up: The study successfully established a of pig skeletal single root muscle fibers in vitro culture methods and satellite cells of high purity; Secondly, the optimization of pig skeletal muscle satellite cell proliferation, differentiation culture system; Finally, a preliminary analysis of the pig skeletal muscle satellite cells into muscle-specific protein expression pattern for the future of skeletal muscle satellite cell migration, proliferation, and differentiation mechanism of providing the ideal experimental material.

Related Dissertations

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Pig
© 2012 www.DissertationTopic.Net Mobile