| Bactrian camel is a special domestic animal with the largest body,the longest life,and the largest living environment.Skeletal muscle is also an important power organ of the body.Bone satellite cells have been successfully established in a variety of animal and human populations through long-term continuous research.The double-bodied experiment is an in-depth understanding of Bactrian camel skeletal muscle satellite cells.The mechanism of skeletal muscle decomposition has potential significance for increasing meat and yield.Using Alxa Bactrian camel skeletal muscle tissue as the sample material,ultra-thin sectioning of the muscle sample was performed using field emission transmission electron microscopy to photograph the muscle sample,and to find skeletal muscle satellite cells between the myometrium and the basement membrane;Tissue block culture method was used to cultivate the primary cells,and the primary skeletal muscle satellite cells were purified in combination with differential adherence.The growth pattern of the satellite cells was determined by growth curve analysis.The transforming gene PAX7 in the skeletal muscle satellite cells was co-opted by immunofluorescence labeling and laser-co-exposed.Focus imaging identification;use PCR amplification method to perform PAX7,MYF5,Desmin gene migration on the sample to obtain more pure Alxa Bactrian camel skeletal muscle satellite cells,so that the satellite cells are located;satellite cells are induced to culture Satellite cells were then subjected to myoblastic differentiation.Grain morphology related research.(1)Conducting transmission electron microscopy studies on the biceps femoris muscle samples of the Bactrian camel from three to June,and skeletal muscle satellite cells were found between the activated and unactivated skeletal muscles between the muscular membrane and the basement membrane of the muscle tissue.The morphology of the satellite cells was observed to be elliptical,with a cross-section of about 5-10um and a width of about 4-7um.The shape of the activated satellite cells is not fixed,and the cell body is incompletely coated.Some cell bodies break through the sarcolemma.There are a lot of electron deposits in the activated satellite cells attached to the cell membrane of the skeletal muscle satellite cells.Compared with the unactivated satellite cells,the shape of the cell membrane is fixed.The internal mitochondria are significantly less than the activated satellite cells and the internal electronic substances are less dispersed.,The two satellite cells have obvious organelles inside the cell body of the wireless mitochondria;(2)Muscle samples were taken from three to six juvenile camels of Bactrian camel,and primary skeletal muscle satellite cells were cultured by two culture methods and compared.The results show that the tissue block culture method is more suitable for the isolation of sterile primary cells after long-distance sampling than the two-step enzymatic cell culture method.Eventually,the recombinant cells are recombined for active recombination,not easy to contaminate,and the culture period is prolonged.to cultivate;(3)Identify the passaged cells by immunofluorescence staining and PCR amplification methods.It was proved that the cell purity reached 80%,and the PAX7,MYF5,and Desmin genes were combined with the corresponding bands after PCR expansion,and finally it was proved that the cultured cells in vitro were skeletal muscle satellite cells with higher purity;(4)Induce differentiation of skeletal muscle satellite cells cultured in the early stage,and use differentiation medium for skeletal muscle satellite cells to induce myogenesis.Satellite cells began to differentiate initially 48 hours after the induction medium was replaced.The cell body was significantly elongated and changed from a short spindle type to a long strip.After one week after the medium was replaced,the satellite cells showed obvious differentiation.Obviously gathered into bundles,obvious myotube structure can be observed. |
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