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不同基础培养基和血清类型组合对原代肉鸡肌肉卫星细胞增殖和分化过程中细胞异质性的影响

Effect of Different Basal Culture Media and Sera Type Combinations on Primary Broiler Chicken Muscle Satellite Cell Heterogeneity during Proliferation and Differentiation.

作者信息

Flees Joshua J, Starkey Charles W, Starkey Jessica D

机构信息

Department of Poultry Science, Auburn University, Auburn, AL 36849, USA.

出版信息

Animals (Basel). 2022 May 31;12(11):1425. doi: 10.3390/ani12111425.

Abstract

The objective of this experiment was to access primary satellite cell (SC) proliferation and differentiation when cultured in different combinations of basal media and sera due to little consistency being published on the optimal culture media for primary broiler chicken satellite cells. Cells were cultured in one of three different basal media: McCoy's 5A, high glucose Dulbecco's Modified Eagle's medium (DMEM), and low glucose DMEM. Media were supplemented with 15% chicken serum (CS) or a combination of 5% horse serum (HS) + 10% CS during proliferation while 3% HS or 3% CS were added to the media during differentiation. Cultures were immunofluorescence stained for myogenic regulatory factors (MRF) at 48, 72, and 96 h post-plating for proliferation (Pax7, MyoD, and Myf-5) and 96 h post-proliferation during differentiation (Pax7 and MyoD), including MF20 to assess fusion. Cells cultured in Dulbecco's Modified Eagle's medium tended to have higher proportions of myogenic cells expressing MRF during proliferation and promoted fusion into myotubes compared with McCoy's 5A during differentiation. Culturing primary SC in low glucose media, glucose concentrations similar to circulating glucose concentrations in broilers, HSCS during proliferation and CS during differentiation, appears to be optimal for promoting broiler chicken satellite cell proliferation and differentiation.

摘要

由于关于原代肉鸡卫星细胞的最佳培养基发表的一致性研究较少,本实验的目的是评估原代卫星细胞(SC)在不同基础培养基和血清组合中培养时的增殖和分化情况。细胞在三种不同的基础培养基之一中培养: McCoy's 5A、高糖杜氏改良 Eagle 培养基(DMEM)和低糖 DMEM。在增殖期间,培养基补充 15%鸡血清(CS)或 5%马血清(HS)+10%CS 的组合,而在分化期间向培养基中添加 3%HS 或 3%CS。在接种后 48、72 和 96 小时对培养物进行免疫荧光染色,以检测增殖期的生肌调节因子(MRF)(Pax7、MyoD 和 Myf-5)以及分化期增殖后 96 小时的(Pax7 和 MyoD),包括 MF20 以评估融合情况。与 McCoy's 5A 相比,在杜氏改良 Eagle 培养基中培养的细胞在增殖期间表达 MRF 的生肌细胞比例往往更高,并且在分化期间促进融合成肌管。在低糖培养基(类似于肉鸡循环葡萄糖浓度的葡萄糖浓度)中培养原代 SC,增殖期使用 HSCS,分化期使用 CS,似乎最有利于促进肉鸡卫星细胞的增殖和分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5812/9179426/f90ea5792c60/animals-12-01425-g001.jpg

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