A Genome-wide Analysis of Human Pluripotent Stem Cell-Derived Endothelial Cells in 2D or 3D Culture.
A defined protocol for efficiently deriving endothelial cells from human pluripotent stem cells was established and vascular morphogenesis was used as a model system to understand how synthetic hydrogels influence global biological function compared with common 2D and 3D culture platforms. RNA sequencing demonstrated that gene expression profiles were similar for endothelial cells and pericytes cocultured in polyethylene glycol (PEG) hydrogels or Matrigel, while monoculture comparisons identified distinct vascular signatures for each cell type. Endothelial cells cultured on tissue-culture polystyrene adopted a proliferative phenotype compared with cells cultured on or encapsulated in PEG hydrogels. The proliferative phenotype correlated to increased FAK-ERK activity, and knockdown or inhibition of ERK signaling reduced proliferation and expression for cell-cycle genes while increasing expression for "3D-like" vasculature development genes. Our results provide insight into the influence of 2D and 3D culture formats on global biological processes that regulate cell function.
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Jue
J
Zhang
Department of Biomedical Engineering, University of Wisconsin-Madison, Wisconsin Institute for Medical Research, Room 5405, Madison, WI 53706, USA. Electronic address: mpschwartz@wisc.edu.
Michael P
MP
Schwartz
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Zhonggang
Z
Hou
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Yongsheng
Y
Bai
Department of Biomedical Engineering, University of Wisconsin-Madison, Wisconsin Institute for Medical Research, Room 5405, Madison, WI 53706, USA.
Hamisha
H
Ardalani
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Scott
S
Swanson
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
John
J
Steill
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Victor
V
Ruotti
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Angela
A
Elwell
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Bao Kim
BK
Nguyen
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Jennifer
J
Bolin
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Ron
R
Stewart
Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA; Department of Cell & Regenerative Biology, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Molecular, Cellular, & Developmental Biology, University of California, Santa Barbara, CA 93117, USA.
James A
JA
Thomson
Department of Biomedical Engineering, University of Wisconsin-Madison, Wisconsin Institute for Medical Research, Room 5405, Madison, WI 53706, USA; Materials Science Program, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Orthopedics and Rehabilitation, University of Wisconsin-Madison, Madison, WI 53705, USA. Electronic address: wlmurphy@wisc.edu.
William L
WL
Murphy
D018929:Cell Culture Techniques / Q000379:methods; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D007797:Laminin / Q000737:chemistry; D020935:MAP Kinase Signaling System; D018919:Neovascularization, Physiologic; D020286:Pericytes / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry; D011509:Proteoglycans / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D059467:Transcriptome
D004338:Drug Combinations; D020100:Hydrogels; D007797:Laminin; D011137:Polystyrenes; D011509:Proteoglycans; C059485:matrigel; D011092:Polyethylene Glycols; D003094:Collagen
3D culture; MAPK; Matrigel; differentiation; endothelial cells; focal adhesion kinase; human pluripotent stem cells; hydrogel; tissue engineering; xeno-free