Mir‑2400 significantly promotes the biological function of bovine skeletal muscle satellite cell proliferation

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

Inactive Publication Date: 2017-09-19
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Mir‑2400 significantly promotes the biological function of bovine skeletal muscle satellite cell proliferation
  • Mir‑2400 significantly promotes the biological function of bovine skeletal muscle satellite cell proliferation
  • Mir‑2400 significantly promotes the biological function of 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

miR-2400 significantly promotes the biological function of the proliferation of bovine skeletal muscle satellite cells, belongs to the field of cell biology technology, and is characterized in that: the bovine skeletal muscle satellite cells are isolated and cultured in vitro, and the differentiation medium containing 2% horse serum is used to induce bovine skeletal muscle satellite cells. Skeletal muscle satellite cell differentiation. After 3 days of induced differentiation, the expression of MHC and Desmin in bovine skeletal muscle satellite cells was observed under a fluorescence microscope; the expression of miR-2400 was detected by stem-loop fluorescent quantitative RT-PCR, and EdU, The effect of miR-2400 on the proliferation of skeletal muscle satellite cells was detected by PCNA immunofluorescence, CCK-8, and the expression of CCND2 and CDK6 by fluorescence quantitative RT-PCR; the above results indicated that miR-2400 could regulate the proliferation and differentiation of bovine skeletal muscle satellite cells It has a certain effect and can significantly promote the proliferation of bovine skeletal muscle satellite cells.

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 Patents(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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