TY - JOUR
T1 - Engineered Microenvironment for Manufacturing Human Pluripotent Stem Cell-Derived Vascular Smooth Muscle Cells
AU - Lin, Haishuang
AU - Qiu, Xuefeng
AU - Du, Qian
AU - Li, Qiang
AU - Wang, Ou
AU - Akert, Leonard
AU - Wang, Zhanqi
AU - Anderson, Dirk
AU - Liu, Kan
AU - Gu, Linxia
AU - Zhang, Chi
AU - Lei, Yuguo
N1 - Funding Information:
This work was partially funded by the UNL-UNMC Science Engineering and Medicine (SEM) initiative grant. Confocal microscope imaging was done in the Morrison Microscopy Core Research Facility at the University of Nebraska, Lincoln. Dr. You Zhou and Dr. Christian Elowsky assisted in the confocal imaging. Flow cytometry was done in the Morrison Center, the Flow Cytometry Service Center, University of Nebraska, Lincoln, with the assistance of Dirk Anderson. RNA sequencing was done at the UNMC deep sequencing core.
Publisher Copyright:
© 2018 The Authors
PY - 2019/1/8
Y1 - 2019/1/8
N2 - Human pluripotent stem cell-derived vascular smooth muscle cells (hPSC-VSMCs) are of great value for disease modeling, drug screening, cell therapies, and tissue engineering. However, producing a high quantity of hPSC-VSMCs with current cell culture technologies remains very challenging. Here, we report a scalable method for manufacturing hPSC-VSMCs in alginate hydrogel microtubes (i.e., AlgTubes), which protect cells from hydrodynamic stresses and limit cell mass to <400 μm to ensure efficient mass transport. The tubes provide cells a friendly microenvironment, leading to extremely high culture efficiency. We have shown that hPSC-VSMCs can be generated in 10 days with high viability, high purity, and high yield (∼5.0 × 10 8 cells/mL). Phenotype and gene expression showed that VSMCs made in AlgTubes and VSMCs made in 2D cultures were similar overall. However, AlgTube-VSMCs had higher expression of genes related to vasculature development and angiogenesis, and 2D-VSMCs had higher expression of genes related to cell death and biosynthetic processes.
AB - Human pluripotent stem cell-derived vascular smooth muscle cells (hPSC-VSMCs) are of great value for disease modeling, drug screening, cell therapies, and tissue engineering. However, producing a high quantity of hPSC-VSMCs with current cell culture technologies remains very challenging. Here, we report a scalable method for manufacturing hPSC-VSMCs in alginate hydrogel microtubes (i.e., AlgTubes), which protect cells from hydrodynamic stresses and limit cell mass to <400 μm to ensure efficient mass transport. The tubes provide cells a friendly microenvironment, leading to extremely high culture efficiency. We have shown that hPSC-VSMCs can be generated in 10 days with high viability, high purity, and high yield (∼5.0 × 10 8 cells/mL). Phenotype and gene expression showed that VSMCs made in AlgTubes and VSMCs made in 2D cultures were similar overall. However, AlgTube-VSMCs had higher expression of genes related to vasculature development and angiogenesis, and 2D-VSMCs had higher expression of genes related to cell death and biosynthetic processes.
UR - http://www.scopus.com/inward/record.url?scp=85059828418&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85059828418&partnerID=8YFLogxK
U2 - 10.1016/j.stemcr.2018.11.009
DO - 10.1016/j.stemcr.2018.11.009
M3 - Article
C2 - 30527760
AN - SCOPUS:85059828418
SN - 2213-6711
VL - 12
SP - 84
EP - 97
JO - Stem Cell Reports
JF - Stem Cell Reports
IS - 1
ER -