Zohora Fatema Tuj, Aldebs Alyaa Isam, Nosoudi Nasim, Singh Surinder Pal, Ramirez-Vick Jaime Eduardo
Department of Biomedical, Industrial, and Human Factors Engineering, Wright State University, Dayton, Ohio, USA.
Biomedical Engineering Program,Marshall University, Huntington, West Virginia, USA.
Cells Tissues Organs. 2019;208(3-4):113-133. doi: 10.1159/000507187. Epub 2020 May 28.
Much of the current understanding on molecular and cellular events of adipose developmental biology comes from monolayer cell culture models using preadipocyte cell lines, although in vivo adipose tissue consists of a much more complex three-dimensional microenvironment of diverse cell types, extracellular network, and tissue-specific morphological and functional features. Added to this fact, the preadipocytes, on which the adipogenesis mechanisms are mostly explored, possess some serious limitations (e.g., time of initial subculture and adipogenic differentiation time), which, perhaps, can efficiently be replaced with progenitor cells such as adipose tissue-derived stem cells (ASCs). With the objective of developing a better in vitro model for adipose developmental biology, this project involves gene expression profiling of human ASCs (hASCs) during their differentiation to adipocytes in a 2D versus 3D culture model. This transcriptional-level analysis revealed that gene expression patterns of adipogenesis-induced hASCs in a 3D self-assembled polypeptide hydrogel are relatively different from the 2D monolayered cells on plastic hard substrate. Moreover, analysis of adipogenic lineage progression 9 days after adipogenic induction shows earlier differentiation of hASCs in 2D over their 3D counterparts. However, differentiation in 2D shows some unexpected behavior in terms of gene expression, which does not seem to be related to adipogenic lineage specification. Since hASCs are already being used in clinical trials due to their therapeutic potential, it is important to have a clear understanding of the molecular mechanisms in an in vivo model microenvironment like the one presented here.
目前对脂肪发育生物学分子和细胞事件的许多理解来自使用前脂肪细胞系的单层细胞培养模型，尽管体内脂肪组织由多种细胞类型、细胞外网络以及组织特异性形态和功能特征组成的更为复杂的三维微环境构成。除此之外，大部分用于探索脂肪生成机制的前脂肪细胞存在一些严重的局限性（例如，首次传代培养时间和成脂分化时间），而这些局限性或许可以用诸如脂肪组织来源干细胞（ASCs）等祖细胞有效地替代。为了开发一个更好的脂肪发育生物学体外模型，本项目涉及在二维与三维培养模型中，对人脂肪组织来源干细胞（hASCs）向脂肪细胞分化过程进行基因表达谱分析。这种转录水平分析表明，在三维自组装多肽水凝胶中脂肪生成诱导的hASCs的基因表达模式与在塑料硬质底物上的二维单层细胞相对不同。此外，对脂肪生成诱导9天后脂肪生成谱系进展的分析表明，二维培养的hASCs比三维培养的hASCs分化更早。然而，二维培养中的分化在基因表达方面表现出一些意外行为，这似乎与脂肪生成谱系的特化无关。鉴于hASCs因其治疗潜力已被用于临床试验，清楚了解此处所呈现的体内模型微环境中的分子机制很重要。
