title: Engineering Strategies for the Formation of Embryoid Bodies from Human Pluripotent Stem Cells.
abstract: Human pluripotent stem cells (hPSCs) are powerful tools for regenerative therapy and studying human developmental biology, attributing to their ability to differentiate into many functional cell types in the body. The main challenge in realizing hPSC potential is to guide their differentiation in a well-controlled manner. One way to control the cell differentiation process is to recapitulate during in vitro culture the key events in embryogenesis to obtain the three developmental germ layers from which all cell types arise. To achieve this goal, many techniques have been tested to obtain a cellular cluster, an embryoid body (EB), from both mouse and hPSCs. Generation of EBs that are homogeneous in size and shape would allow directed hPSC differentiation into desired cell types in a more synchronous manner and define the roles of cell-cell interaction and spatial organization in lineage specification in a setting similar to in vivo embryonic development. However, previous success in uniform EB formation from mouse PSCs cannot be extrapolated to hPSCs possibly due to the destabilization of adherens junctions on cell surfaces during the dissociation into single cells, making hPSCs extremely vulnerable to cell death. Recently, new advances have emerged to form uniform human embryoid bodies (hEBs) from dissociated single cells of hPSCs. In this review, the existing methods for hEB production from hPSCs and the results on the downstream differentiation of the hEBs are described with emphases on the efficiency, homogeneity, scalability, and reproducibility of the hEB formation process and the yield in terminal differentiation. New trends in hEB production and directed differentiation are discussed.
authors.affiliation: 1 Department of Biomedical Engineering, Virginia Commonwealth University , Richmond, Virginia.|2 Department of Chemical and Life Science Engineering, Virginia Commonwealth University , Richmond, Virginia.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Pettinato
authors.affiliation: 2 Department of Chemical and Life Science Engineering, Virginia Commonwealth University , Richmond, Virginia.|3 Shanghai East Hospital, The Institute for Biomedical Engineering and Nano Science, Tongji University School of Medicine, Shanghai, People's Republic of China .
authors.forename: Xuejun
authors.initials: X
authors.lastname: Wen
authors.affiliation: 1 Department of Biomedical Engineering, Virginia Commonwealth University , Richmond, Virginia.
authors.forename: Ning
authors.initials: N
authors.lastname: Zhang
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology*; D047108:Embryonic Development / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice