Laboratory of Stem Cell Biology and Engineering, Department of Animal Sciences, University of Illinois at Urbana-Champaign, Urbana, Ill., USA.
Cell Tissue Res. 2010 Sep;341(3):359-70. doi: 10.1007/s00441-010-1015-3. Epub 2010 Aug 3.
Advances in bioengineering, material chemistry, and developmental biology have led to the design of three-dimensional (3D) culture systems that better resemble the surrounding structure and chemistry of the in situ niches of cells in tissues. This study was designed to characterize and compare porcine adipose-derived stem cells (ADSC) and bone-marrow-derived stem cells (BMSC) induced to differentiate toward osteogenic and adipogenic lineages in vitro by using a 3D alginate hydrogel. The morphology and gene expression of the two cell populations during differentiation were analyzed. Both ADSC and BMSC showed morphological evidence of osteogenic and adipogenic differentiation. Expression patterns of genes characteristic of the onset of osteogenic differentiation (ALP, COL1A1, SPARC, SPP1) were low at the beginning of culture and generally increased during the period of differentiation up to 28 days in culture. Expression of genes associated with adipogenic differentiation (ACSL1, ADFP, ADIPOQ, CD36, DBI, DGAT2, PPARG, SCD) was consistently increased in ADSC cultured in alginate hydrogel relative to the start of differentiation. However, adipogenic gene expression of BMSC cultured in alginate hydrogel was more limited when compared with that of ADSC. Evaluation of cell numbers (via the MTT staining assay) suggested a greater viability of BMSC under osteogenic conditions in alginate hydrogels than under adipogenic conditions, whereas ADSC had greater viability under adipogenic conditions than under osteogenic conditions. This study thus provides an important initial evaluation of ADSC and BMSC seeded and differentiated toward the osteogenic and adipogenic cell lineages in a 3D alginate hydrogel in vitro.
生物工程、材料化学和发育生物学的进步促使人们设计出了三维（3D）培养系统，这些系统更能模拟细胞在组织原位微环境中的结构和化学特性。本研究旨在利用 3D 藻酸盐水凝胶，对体外诱导向成骨和成脂谱系分化的猪脂肪来源干细胞（ADSC）和骨髓来源干细胞（BMSC）进行鉴定和比较。分析了两种细胞群在分化过程中的形态和基因表达。ADSC 和 BMSC 均表现出向成骨和成脂分化的形态学证据。成骨分化起始特征基因（ALP、COL1A1、SPARC、SPP1）的表达模式在培养开始时较低，通常在分化过程中增加，直到培养 28 天。与分化开始时相比，在藻酸盐水凝胶中培养的 ADSC 中，与脂肪生成分化相关的基因（ACSL1、ADFP、ADIPOQ、CD36、DBI、DGAT2、PPARG、SCD）的表达持续增加。然而，与 ADSC 相比，在藻酸盐水凝胶中培养的 BMSC 的脂肪生成基因表达更为有限。通过 MTT 染色法评估细胞数量表明，与脂肪生成条件相比，BMSC 在藻酸盐水凝胶中成骨条件下具有更高的存活率，而 ADSC 在脂肪生成条件下的存活率高于成骨条件下。因此，本研究首次对在 3D 藻酸盐水凝胶中体外向成骨和成脂细胞谱系接种和分化的 ADSC 和 BMSC 进行了重要评估。
