Protocol-Dependent Morphological Changes in Human Embryonic Stem Cell Aggregates during Differentiation toward Early Pancreatic Fate.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
ACS Applied Bio Materials Pub Date : 2024-01-01 Epub Date: 2022-11-14 DOI:10.1159/000527863
Elmira Rezaei Zonooz, Zahra Ghezelayagh, Azadeh Moradmand, Hossein Baharvand, Yaser Tahamtani
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Abstract

Cell therapy is one of the promising approaches used against type 1 diabetes. Efficient generation of human embryonic stem cell (hESC)-derived pancreatic progenitors (PPs) is of great importance. Since signaling pathways underlying human pancreas development are not yet fully understood, various differentiation protocols are conducted, each considering variable duration, timing, and concentrations of growth factors and small molecules. Therefore, we compared two PP differentiation protocols in static suspension culture. We tested modified protocols developed by Pagliuca et al. (protocol 1) and Royan researchers (protocol 2) until early PP stage. The morphological changes of hESC aggregates during differentiation, and also gene and protein expression after differentiation, were evaluated. Different morphological structures were formed in each protocol. Quantitative gene expression analysis, flow cytometry, and immunostaining revealed a high level of PDX1 expression on day 13 of Royan's differentiation protocol compared to protocol 1. Our data showed that using protocol 2, cells were further differentiated until day 16, showing higher efficiency of early PPs. Moreover, protocol 2 is able to produce hESCs-PPs in a static suspension culture. Since protocol 2 is inexpensive in terms of media, growth factors, and chemicals, it can be used for massive production of PPs using static and dynamic suspension cultures.

人类胚胎干细胞聚集体在向早期胰腺命运分化过程中发生的形态学变化。
细胞疗法是治疗1型糖尿病的有效方法之一。高效生成人类胚胎干细胞(hESC)衍生的胰腺祖细胞(PPs)非常重要。由于人类胰腺发育的信号通路尚未完全明了,因此进行了各种分化方案,每种方案都考虑了不同的持续时间、时机以及生长因子和小分子的浓度。因此,我们比较了静态悬浮培养中的两种胰腺分化方案。我们测试了 Pagliuca 等人(方案-1)和 Royan 研究人员(方案-2)开发的改良方案,直至 PP 早期阶段。我们对分化过程中 hESC 聚集体的形态变化以及分化后的基因和蛋白质表达进行了评估。每个方案都形成了不同的形态结构。定量基因表达分析、流式细胞术和免疫染色显示,与方案-1相比,Royan分化方案第13天的PDX1表达水平较高。我们的数据显示,使用方案-2,细胞可进一步分化至第 16 天,显示早期 PPs 的效率更高。此外,方案-2 还能在静态悬浮培养中产生 hESCs-PPs。由于方案-2 在培养基、生长因子和化学物质方面成本低廉,因此可用于利用静态和动态悬浮培养大规模生产 PPs。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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