Substrate stiffness effect and chromosome missegregation in hIPS cells
Substrate stiffness effect and chromosome missegregation in hIPS cells
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DOI：
10.1186/s12952-015-0042-8
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发表时间：
2015-01-01
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影响因子：
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通讯作者：
Crone, Wendy C.
中科院分区：
文献类型：
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作者：
Acevedo-Acevedo, Suehelay;Crone, Wendy C.
背景：确保多能干细胞（PSC）培养物的遗传稳定性对于成功开发细胞疗法至关重要。虽然大多数情况导致实验失败，文献中没有报道，但许多实验室发现，在体外培养长时间后，psc中出现了遗传异常。这些细胞主要在非生理性硬基质中培养，如组织培养聚苯乙烯（TCPS），这增加了这些异常的原因可能受到基质力学影响的可能性。研究结果：为了研究这一点，人类psc在不同硬度的基质上生长，如一系列聚丙烯酰胺配方、TCPS和硼硅酸盐玻璃盖。这些衬底允许测试从5kPa到64GPa的刚度范围。本研究分析了两种人诱导PSC （iPSC）系：19-9-11 iPSC和19.7克隆F iPSC。中心体和DNA染色显示，在不同的培养条件下，19-9-11 iPSCs的有丝分裂率在1- 8.5%之间。19.7个克隆的fipscs有丝分裂异常发生率为4.4% ~ 8.1%。结论：异常细胞分裂不偏向于一种特定的底物。方差分析（ANOVA）和Tukey’s Honest Significant Difference检验证实传代数、细胞系或底物之间没有统计学上的显著差异。
Background: Ensuring genetic stability in pluripotent stem cell (PSC) cultures is essential for the development of successful cell therapies. Although most instances lead to failed experiments and go unreported in the literature, many laboratories have found the emergence of genetic abnormalities in PSCs when cultured in vitro for prolonged amounts of time. These cells are primarily cultured in non-physiological stiff substrates like tissue culture polystyrene (TCPS) which raises the possibility that the cause of these abnormalities may be influenced by substrate mechanics.Findings: In order to investigate this, human PSCs were grown on substrates of varying stiffness such as a range of polyacrylamide formulations, TCPS, and borosilicate glass coverslips. These substrates allowed for the testing of a stiffness range from 5kPa to 64GPa. Two human induced PSC (iPSC) lines were analyzed in this study: 19-9-11 iPSCs and 19.7 clone F iPSCs. Centrosome and DNA staining revealed that 19-9-11 iPSCs range from 1-8.5 % abnormal mitoses under the different culture conditions. A range of 4.4-8.1 % abnormal mitoses was found for 19.7 clone F iPSCs.Conclusions: Abnormal cell division was not biased towards one particular substrate. It was confirmed by Analysis of Variance (ANOVA) and Tukey's Honest Significant Difference test that there was no statistically significant difference between passage numbers, cell lines, or substrates.