Cyclic Strain Promotes H19 Expression and Vascular Tube Formation in iPSC-Derived Endothelial Cells
- PDF / 1,415,181 Bytes
- 9 Pages / 593.972 x 792 pts Page_size
- 52 Downloads / 175 Views
Cellular and Molecular Bioengineering ( 2020) https://doi.org/10.1007/s12195-020-00617-0
Original Article
Cyclic Strain Promotes H19 Expression and Vascular Tube Formation in iPSC-Derived Endothelial Cells MARK J. VANDER ROEST and W. DAVID MERRYMAN Biomedical Engineering, Vanderbilt University, Room 9445D MRB4, 2213 Garland Ave, Nashville, TN 37212, USA (Received 23 October 2019; accepted 28 April 2020) Associate Editor Michael R. King oversaw the review of this article.
Abstract Introduction—Induced pluripotent stem cell (iPSC)-derived endothelial cells (ECs) have the potential for therapeutic application in several cardiovascular diseases. Mechanical strain is known to regulate EC behavior and stem cell differentiation and may play a role in directing EC differentiation of iPSCs. H19, a long non-coding RNA (lncRNA), is known to affect ECs in several mechanically relevant pathologies and may play a role in this process as well. Therefore, we investigated expression changes of H19 resulting from mechanical stimulation during EC differentiation, as well as functional effects on EC tube formation. Methods—iPSCs were subjected to 5% cyclic mechanical strain during EC differentiation. RT-PCR and flow cytometry were used to assess changes in mesoderm differentiation and gene expression in the final ECs as a result of strain. Functional outcomes of mechanically differentiated ECs were assessed with a tube formation assay and changes in H19. H19 was also overexpressed in human umbilical vein endothelial cells (HUVECs) to assess its role in non-H19expressing ECs. Results—Mechanical strain promoted mesoderm differentiation, marked by increased expression of brachyury 24 h after initiation of differentiation. Strain also increased expression of H19, CD31, VE-cadherin, and VEGFR2 in differentiated ECs. Strain-differentiated ECs formed tube networks with higher junction and endpoint density than statically-differentiated ECs. Overexpression of H19 in HUVECs resulted in similar patterns of tube formation. Conclusions—H19 expression is increased by mechanical strain and promotes tube branching in iPSC-derived ECs. Keywords—lncRNA, Angiogenesis, Mechanobiology, Stem cells.
Address correspondence to W. David Merryman, Biomedical Engineering, Vanderbilt University, Room 9445D MRB4, 2213 Garland Ave, Nashville, TN 37212, USA. Electronic mail: [email protected]
INTRODUCTION Endothelial cells (ECs) line the interior of the circulatory system and are critically involved in a variety of cardiovascular functions and diseases. During healthy function, ECs regulate nutrient and oxygen exchange, mediate immune cell infiltration from the circulatory system to target tissues, maintain vascular tone, and are crucial for the formation of new blood vessels.24 The endothelium is also directly involved in numerous cardiovascular diseases, such as atherosclerosis, coronary artery disease, or valvular heart diseases, and induced pluripotent stem cell (iPSC)-derived ECs are increasingly being considered for cell therapy approaches to r
Data Loading...
