Method using a co-culture system with high-purity intramuscular preadipocytes and satellite cells from chicken pectoralis major muscle

文献类型： 外文期刊

第一作者： Cui, H. X.

作者： Cui, H. X.;Guo, L. P.;Zhao, G. P.;Liu, R. R.;Li, Q. H.;Zheng, M. Q.;Wen, J.;Cui, H. X.;Guo, L. P.;Zhao, G. P.;Liu, R. R.;Li, Q. H.;Zheng, M. Q.;Wen, J.;Guo, L. P.

作者机构：

关键词： Chicken; new method; intramuscular preadipocyte isolation; muscle satellite cell; co-culture

期刊名称：POULTRY SCIENCE （ 影响因子：3.352； 五年影响因子：3.679 ）

ISSN： 0032-5791

年卷期： 2018 年 97 卷 10 期

页码：

收录情况： SCI

摘要： Intramuscular fat is important in improving meat quality; however, the lack of high-purity intramuscular preadipocyte (IMP) in vitro has severely limited the in-depth research on the mutual regulation of myocytes and adipocytes in chicken. In this study, we establish a new method by combining the mature adipocyte ceiling method and the transwell co-culture system. Mature intramuscular adipocyte (MIA) and muscle satellite cell (MSC) were obtained from digested pectoralis major, and MIAs were transformed into IMPs by dedifferentiation with ceiling culture. MSCs were then purified by differential adhesion for 2 h. The results by inverted-microscope observation, MTT assay, Oil Red O staining, and q-PCR revealed that the de-differentiated cells from MIA were identified as the IMPs, and had the same the cellular morphology, the capacity on differentiation, proliferation and passage with the abdominal fat preadipocytes (P > 0.05). The applicability of the obtained IMPs in co-cultured experiment with the MSC revealed that it could meet the requirements of the experimental study. Finally, a co-culture system of IMPs and MSCs was established using a transwell chamber. The co-cultured results indicated that MSCs in the proliferative stage tend to accelerate the differentiation of IMPs to induce more fat content in co-cultured IMPs than in single-culture IMPs (P < 0.05), in the non-proliferative stage, the results tend to show the opposite (P < 0.05). The mRNA levels of related genes significantly changed in accordance with the fat content in cells. The results strongly supported the view that the established co-culture system was effective and feasible. In summary, we successfully found a new method to explore the interaction between myocytes and adipocytes of chicken. Our findings can deepen research on the regulation of chicken myocytes and adipocytes.

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