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Establishment and validation of cell pools using primary muscle cells derived from satellite cells of pig skeletal muscle.
In Vitro Cellular & Developmental Biology - Animal ( IF 2.1 ) Pub Date : 2019-12-23 , DOI: 10.1007/s11626-019-00428-2
Katharina Metzger 1, 2 , Armin Tuchscherer 3 , Marie-France Palin 4 , Siriluck Ponsuksili 2 , Claudia Kalbe 1
Affiliation  

Primary cell cultures derived from satellite cells of skeletal muscle provide an appropriate in vitro model for proliferating myoblasts and differentiating myotubes for muscle biological research. These cell cultures may consist of harvested cells per animal or of a cell pool made of cells from several animals. However, cell pooling reduces the biological variability of the different cell donors. On the other hand, the use of cell pools offers an opportunity to use less donor tissue and to perform long-term projects with a broad spectrum of analysis and replications. In the literature, information about the donors of cell pools, the procedure used for pooling, and the characterization/validation of cell pools is often lacking. In this study, we established three cell pools consisting of M. rhomboideus or M. longissimus from ten or six piglets, each with one gender and medium birth weight. Real-time impedimetric monitoring was used to evaluate the proliferative growth behavior of myoblasts for the cell pools in comparison to their corresponding unpooled cells over a period of 72 h, with a measurement being taken every 30 min. For each of the tested cell pools, cell index, slope, and doubling time did not differ between the cell pool and the unpooled cells of the donor animals. Differentiation capacity and mRNA expression of PAX7, MYOD and MYOG remained unchanged between the cell pool and the unpooled cells. Current results support that the use of cell pools is an appropriate method to reflect the average proliferative growth behavior of unpooled cells.

中文翻译:

使用源自猪骨骼肌卫星细胞的原代肌肉细胞建立和验证细胞池。

源自骨骼肌卫星细胞的原代细胞培养物为增殖成肌细胞和分化肌管提供了合适的体外模型,以进行肌肉生物学研究。这些细胞培养物可由每只动物收获的细胞组成,或由数只动物的细胞组成的细胞池组成。但是,细胞合并降低了不同细胞供体的生物学变异性。另一方面,细胞池的使用提供了一个机会,可以使用较少的供体组织,并进行具有广泛分析和复制范围的长期项目。在文献中,通常缺乏有关细胞池供体,用于池化的程序以及细胞池表征/验证的信息。在这项研究中,我们建立了三个细胞池,分别由十个或六个仔猪的菱形支原体或长支原体组成,每个人的性别和出生体重中等。实时阻抗监测用于评估成肌细胞在72小时内与其对应的非池化细胞相比在细胞池中的增殖生长行为,每30分钟进行一次测量。对于每个测试的细胞池,细胞池,斜率和倍增时间在细胞池和供体动物的未池化细胞之间没有差异。PAX7,MYOD和MYOG的分化能力和mRNA表达在细胞池和未池化细胞之间保持不变。目前的结果支持使用细胞池是反映未池化细胞的平均增殖生长行为的合适方法。实时阻抗监测用于评估成肌细胞在72小时内与其对应的非池化细胞相比在细胞池中的增殖生长行为,每30分钟进行一次测量。对于每个测试的细胞池,细胞池,斜率和倍增时间在细胞池和供体动物的未池化细胞之间没有差异。PAX7,MYOD和MYOG的分化能力和mRNA表达在细胞池和未池化细胞之间保持不变。目前的结果支持使用细胞池是反映未池化细胞的平均增殖生长行为的合适方法。实时阻抗监测用于评估成肌细胞在72小时内与其对应的非池化细胞相比在细胞池中的增殖生长行为,每30分钟进行一次测量。对于每个测试的细胞池,细胞池,斜率和倍增时间在细胞池和供体动物的未池化细胞之间没有差异。PAX7,MYOD和MYOG的分化能力和mRNA表达在细胞池和未池化细胞之间保持不变。目前的结果支持使用细胞池是反映未池化细胞的平均增殖生长行为的合适方法。细胞池,斜率和倍增时间在供体动物的细胞池和未池化细胞之间没有差异。PAX7,MYOD和MYOG的分化能力和mRNA表达在细胞池和未池化细胞之间保持不变。目前的结果支持使用细胞池是反映未池化细胞的平均增殖生长行为的合适方法。细胞池,斜率和倍增时间在供体动物的细胞池和未池化细胞之间没有差异。PAX7,MYOD和MYOG的分化能力和mRNA表达在细胞池和未池化细胞之间保持不变。目前的结果支持使用细胞池是反映未池化细胞的平均增殖生长行为的合适方法。
更新日期:2019-12-23
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