Mossakowski Medical Research Centre, Translational Platform for Regenerative Medicine, Polish Academy of Science, 02-106 Warsaw, Poland.
Mossakowski Medical Research Centre, Department of Stem Cell Bioengineering, Polish Academy of Sciences, 02-106 Warsaw, Poland.
Cells. 2021 Mar 24;10(4):719. doi: 10.3390/cells10040719.
The aim of our work was to develop a protocol enabling a derivation of mesenchymal stem/stromal cell (MSC) subpopulation with increased expression of pluripotent and neural genes. For this purpose we used a 3D spheroid culture system optimal for neural stem cells propagation. Although 2D culture conditions are typical and characteristic for MSC, under special treatment these cells can be cultured for a short time in 3D conditions. We examined the effects of prolonged 3D spheroid culture on MSC in hope to select cells with primitive features. Wharton Jelly derived MSC (WJ-MSC) were cultured in 3D neurosphere induction medium for about 20 days in vitro. Then, cells were transported to 2D conditions and confront to the initial population and population constantly cultured in 2D. 3D spheroids culture of WJ-MSC resulted in increased senescence, decreased stemness and proliferation. However long-termed 3D spheroid culture allowed for selection of cells exhibiting increased expression of early neural and SSEA4 markers what might indicate the survival of cell subpopulation with unique features.
我们的工作旨在开发一种方案，以增加多能性和神经基因的表达来衍生间充质干细胞（MSC）亚群。为此，我们使用了最适合神经干细胞增殖的 3D 球体培养系统。尽管 2D 培养条件是 MSC 的典型特征，但在特殊处理下，这些细胞可以在 3D 条件下短时间培养。我们研究了延长 3D 球体培养对 MSC 的影响，希望选择具有原始特征的细胞。从 Wharton 果冻中分离的 MSC（WJ-MSC）在 3D 神经球诱导培养基中体外培养约 20 天。然后，细胞被转移到 2D 条件下，并与初始群体和不断在 2D 中培养的群体进行对抗。WJ-MSC 的 3D 球体培养导致衰老增加、干性和增殖减少。然而，长期的 3D 球体培养允许选择表达早期神经和 SSEA4 标志物的细胞，这可能表明具有独特特征的细胞亚群的存活。
