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Biological function of miR-2400 in remarkably promoting bovine skeletal muscle satellite cell proliferation

A technology of mir-2400, 1. mir-2400, applied in the field of cell biology, can solve problems such as no biological function reports

Inactive Publication Date: 2015-09-02
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Description
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Problems solved by technology

The literature found miR-2400, but did not report its biological function

Method used

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  • Biological function of miR-2400 in remarkably promoting bovine skeletal muscle satellite cell proliferation
  • Biological function of miR-2400 in remarkably promoting bovine skeletal muscle satellite cell proliferation
  • Biological function of miR-2400 in remarkably promoting bovine skeletal muscle satellite cell proliferation

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Embodiment 1

[0040] 1. The specific steps of the in vitro isolation and culture of bovine skeletal muscle satellite cells: (1) Get 5-10 grams of skeletal muscle tissue from the leg of a newborn calf, wash it with PBS phosphate buffer saline, and cut the tissue into minced shape (about 1 cubic millimeter in size) ), PBS phosphate buffered saline repeatedly pipetting and standing for 1 minute. Aspirate the floating tissue, and centrifuge at 1000 rpm for 10 minutes to obtain myometrium sediment. (2) Add 50 ml of 0.2% type I collagenase to the tissue sediment, and gently shake (about 100 rpm) in a water-bath shaker at 37°C for 2 hours to digest. Then add 40 ml of PBS phosphate buffer solution to the digestion solution to dilute the type I collagenase digestion solution, blow it evenly, transfer it to a centrifuge tube, and centrifuge at 1000 rpm for 10 minutes to remove the type I collagenase digestion solution and pour into the precipitate Add 20 ml of 0.1% trypsin to the solution, and diges...

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Abstract

The invention discloses a biological function of miR-2400 in remarkably promoting bovine skeletal muscle satellite cell proliferation and belongs to the technical field of cell biology. The biological function is characterized in that bovine skeletal muscle satellite cells are subjected to in-vitro separation and cultivation, a differential medium containing 2% horse serum is adopted to induce differentiation of the bovine skeletal muscle satellite cells, and after induced differentiation for 3 days, expression condition of MHC and Desmin in the bovine skeletal muscle satellite cells can be observed under a fluorescent microscope; stem-loop fluorescence quantitative RT-PCR is adopted to detect expression quantity of miR-2400, and fluorescence quantitative RT-PCR of EdU, PCNA immune fluorescence, CCK-8 and CCND2 and CDK6 expression quantity is adopted to detect influence, on bovine skeletal muscle satellite cell proliferation, of miR-2400; the above result shows that miR-2400 has certain effect on adjusting proliferation and differentiation of the bovine skeletal muscle satellite cells and can remarkably promote bovine skeletal muscle satellite cell proliferation.

Description

technical field [0001] The invention relates to the biological function that miR-2400 significantly promotes the proliferation of bovine skeletal muscle satellite cells, and belongs to the technical field of cell biology. Background technique [0002] Skeletal muscle satellite cells are precursor cells of muscle tissue with strong plasticity and many important biological characteristics of skeletal muscle. The activation, proliferation and differentiation of muscle satellite cells are also important mechanisms for skeletal muscle cell hypertrophy, increased number and muscle fiber transformation. Skeletal muscle The proliferation and differentiation process of satellite cells is very complex, involving the expression of a large number of genes and network regulation. Studies have confirmed that miRNA plays a key regulatory role in muscle development and muscle cell proliferation and differentiation (Luo et al., 2013). Therefore, the study of the molecular mechanism of miRNA...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
Inventor 严云勤张伟伟佟慧丽李树峰
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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