title: ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
abstract: We report a technology to form human embryoid bodies (hEBs) from singularized human embryonic stem cells (hESCs) without the use of the p160 rho-associated coiled-coil kinase inhibitor (ROCKi) or centrifugation (spin). hEB formation was tested under four conditions: +ROCKi/+spin, +ROCKi/-spin, -ROCKi/+spin, and -ROCKi/-spin. Cell suspensions of BG01V/hOG and H9 hESC lines were pipetted into non-adherent hydrogel substrates containing defined microwell arrays. hEBs of consistent size and spherical geometry can be formed in each of the four conditions, including the -ROCKi/-spin condition. The hEBs formed under the -ROCKi/-spin condition differentiated to develop the three embryonic germ layers and tissues derived from each of the germ layers. This simplified hEB production technique offers homogeneity in hEB size and shape to support synchronous differentiation, elimination of the ROCKi xeno-factor and rate-limiting centrifugation treatment, and low-cost scalability, which will directly support automated, large-scale production of hEBs and hESC-derived cells needed for clinical, research, or therapeutic applications.
authors.affiliation: Department of Chemical and Life Science Engineering, Virginia Commonwealth University, Richmond, Virginia, United States of America; Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, Virginia, United States of America; Department of Bioengineering, Clemson University, Clemson, South Carolina, United States of America.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Pettinato
authors.affiliation: Department of Chemical and Life Science Engineering, Virginia Commonwealth University, Richmond, Virginia, United States of America; Department of Bioengineering, Clemson University, Clemson, South Carolina, United States of America; Department of Craniofacial Biology, Medical University of South Carolina, Charleston, South Carolina, United States of America.
authors.forename: Wendy S
authors.initials: WS
authors.lastname: Vanden Berg-Foels
authors.affiliation: Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, Virginia, United States of America; Department of Bioengineering, Clemson University, Clemson, South Carolina, United States of America.
authors.forename: Ning
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Department of Chemical and Life Science Engineering, Virginia Commonwealth University, Richmond, Virginia, United States of America; Department of Bioengineering, Clemson University, Clemson, South Carolina, United States of America; Department of Craniofacial Biology, Medical University of South Carolina, Charleston, South Carolina, United States of America; Institute for Biomedical Engineering and Nano Science (iNANO), Shanghai East Hospital, Tongji Medical School, Tongji University, Shanghai, People's Republic of China.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Wen
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D058732:Embryoid Bodies / Q000187:drug effects / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D005855:Germ Layers / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D006801:Humans; D038081:Organogenesis; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D047428:Protein Kinase Inhibitors; D054460:rho-Associated Kinases