Ultrastructural analysis of human umbilical cord derived MSCs at undifferentiated stage and during osteogenic and adipogenic differentiation

Abstract

Mesenchymal stem cells (MSCs) are considered as an important tool for regenerative medicine and experimental treatments. Unveiling the ultrastructural changes during the differentiation of MSCs might help us to understand the nature of the process and to develop novel therapeutic approaches. For this purpose, human umbilical cord (hUC) was chosen as MSC source. In the first place, MSCs were isolated from sub-amniotic, intervascular and perivascular areas of hUC by enzymatic and tissue explant method to determine the most favorable region of hUC and technique for further processing. Therefore, microscopic and growth kinetics analyses showed that there was no clear difference in the morphologies and proliferation rates among the hUC-MSC groups. Flow cytometric analysis showed that CD44 and CD90 MSC markers were highly expressed, while CD34 and CD45 hematopoietic stem cells markers were expressed at low degree. Because our preliminary results showed that there was no conspicuous superiority among the hUC-MSCs groups, whole UC was utilized as a source, and tissue explant method was applied to isolate MSCs for further differentiation analysis. At the 1 and 3 week of osteogenic and adipogenic differentiation, ultrastructural analysis showed an increase in the number of secondary lysosomes in comparison with the undifferentiated status. Increase in the mitochondrial content was also detected at the 1 week of adipogenic differentiation. Consequently, ultrastructural changes including increase in the number of mitochondria and secondary lysosomes during the adipogenic and osteogenic differentiation could be attributed to the switch in energy metabolism of the MSCs and increment in the lysosomal activity respectively.

Efficacy of resveratrol in the wound healing process by reducing oxidative stress and promoting fibroblast cell proliferation and migration

“…The spheroids established by 3T3 fibroblasts from the final experimental groups were prepared and selected for the TEM analysis, in accordance with the literature. 21 Araldite blocks were trimmed, semi-thin sections were analyzed, and finally, ultra-thin sections were contrasted for evaluation and photographed under the TEM (Jeol: JEM-1011). On the one hand, the control and 1 μM RES groups showed statistically similar results for the PDT index ( Figure 1A).…”

Photobiomodulation Therapy in the Proliferation and Differentiation of Human Umbilical Cord Mesenchymal Stem Cells: An In Vitro Study

“…21 Thus, these cell responses are believed to occur through the synthesis of ATP, the increased potential of the mitochondrial membranes and the levels of cyclic adenosine monophosphate. [22][23][24] In this regard, in order to use PBMT, it is important to establish the parameters of irradiation since different standards may produce different effects, especially regarding cell proliferation, 25,26 inducing stimulatory or inhibitory responses. 27,28 Indeed, when we compared the irradiation parameters used, we observed ample divergence between these variables, demonstrating the lack of protocol standardization and the consequent modification of the results.…”

The Effect of miR122 and miR185 on Hepatic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells

“…The pluripotent stem cells surface markers express in hUC-MSCs including Oct-4, Sox-2 [36] . hUC-MSCs have the potential of multidirectional differentiation, and which can differentiate into chondrocytes [37] , hepatocytes [38] , and adipocytes [39] . The cells we obtained from umbilical cord express CD90 (99.16% ± 0.85%) and CD105 (99.26% ± 0.78%), but no expression of CD34 (0.59% ± 0.16%) and CD45 (1.50% ± 2.04%) by these cells and 66.57% ± 4.61% cells expressed positively Oct-4 analyzed by ow cytometry.…”