(176i) Differentiating Human Pluripotent Stem Cells into Vascular Smooth Muscle Cells in Three Dimensional Thermoreversible Hydrogels

Wang, O., University of Nebraska-Lincoln
Lin, H., University of Nebraska, Lincoln
Li, Q., University of Nebraska, Lincoln
Du, Q., University of Nebraska-Lincoln
Duan, B., University of Nebraska Medical Center
Lei, Y. L., University of Nebraska - Lincoln
Zhang, C., University of Nebraska-Lincoln

letter-spacing:-.25pt;background:transparent">Vascular smooth muscle cells
(VSMCs) are of great value and are needed in large quantities for tissue
engineering, drug screening, disease modeling and cell-based therapies.
However, getting high quantity VSMCs remains a challenge. Here, we report a
method for the scalable manufacturing of VSMCs from human pluripotent stem
cells (hPSCs). hPSCs are expanded and differentiated into VSMCs in a three
dimensional (3D) thermoreversible hydrogel. The hydrogel not only acts as a 3D
scaffold for cells to grow, but also protects cells from hydrodynamic stresses
in the culture vessel and prevents cells from excessive aggregation. Together,
the hydrogel creates a cell-friendly microenvironment, leading to high culture
efficiency. We show that VSMCs can be generated in 10 days with high viability
(>90%), high purity (>80%) and high yield (_ line-height:107%;font-family:Arial;letter-spacing:-.25pt;background:transparent">2.0
¡Á 107 cells per mL hydrogel) in the hydrogel scaffold. The
generated VSMCs have normal functions. Genome-wide gene expression analysis
shows VSMCs made in the hydrogel (i.e. 3D-VSMCs)
have higher expression of genes related to vasculature development and
glycolysis compared to VSMCs made in the conventional 2D cultures (2D-VSMCs),
while 2D-VSMCs have higher expression of genes related to cell proliferation.
This simple, defined and efficient method is scalable for manufacturing
hPSC-VSMCs for various biomedical applications. (Published in Biomaterials
. 2019;7(1):347-61.