MD, 1972 - Hebrew University, Israel
DSc, 1982 - Technion, Israel
Development of platform technologies and protocols for directed differentiation of human pluripotent stem cells
Human pluripotent stem cells (embryonic and induced) are capable of becoming cells of all three embryo germ layers, generating each and every tissue of the human body. This unique quality underlies our research focus. By optimizing and controlling the cell's growth parameters, we not only gain a better understanding of early normal and abnormal developmental events, but also are able to direct the cells to commit to any desired cell type. Currently there are five complementary approaches being taken towards developing platform technologies for the culture and scale-up of pluripotent stem cells. First, methods and protocols for the culture of undifferentiated human pluripotent stem cells in animal-, serum- and feeder-layer free conditions are being devised that will be suitable for future industrial and clinical applications. Second, new cell lines (embryonic and induced) are being derived in animal- and feeder-layer-free culture conditions in line with good laboratory practices (GLP). Third, existing human pluripotent stem cells are being characterized using genomics and proteomics methods to expand our knowledge about the features of these lines. Fourth, undifferentiated human pluripotent stem cells are being expanded before differentiation in dynamic cell culture (bioreactors); the goal is to devise a scalable protocol suitable for mass production of desired cell types. Fifth, human pluripotent stem cells are being grown and differentiated in 3D structures as a basis for tissue engineering. In parallel, various approaches are being taken towards developing protocols for the directed differentiation of human pluripotent stem cells. The focus is generation of vasculogenic derivatives, including endothelial cells, multi-lineage pericytes and human mesenchymal stem cells.
(Upper panel) Human blood vessel composed of endothelial cells (red) and pericytes (green) engineered from human pluripotent stem cells using a Matrigel implant. Nuclear staining by DAPI (blue). (Lower Panel) (A) Ectopic bone formation (black circle) in vivo. Inset: massive mineralization of bone matrix positively stained for Alizarin red. (B) Immunofluorescence staining of implants reveals extensive expression of osteopontin (green). Positive labeling with human specific MHC class I antibodies (red) as well as in situ hybridization for human chromosome 1 (inset, red), confirms human identity. Nuclear staining by DAPI (blue). (C) Extensive network of blood vessels containing erythrocytes (arrows) invaded into the implant.
Domev H, Amit M, Dar A, Itskovitz-Eldor J. Efficient Engineering of Vascularized Ectopic Bone from Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells. Tiss Eng. 2012 (in press)
Dar A, Domev H, Ben-Yosef O, Tzukerman M, Zeevi-Levin N, Novak A, Germanguz I, Amit M, Itskovitz-Eldor J. 2012. Multipotent vasculogenic pericytes from human pluripotent stem cells promote recovery of murine ischemic limb. Circulation 125, 87-99.
Amit M, Chebath J, Marguletz V, Laevsky I, Miropolsky Y, Shariki K, Peri M, Revel M, Itskovitz-Eldor J. 2010. Suspension culture of undifferentiated human embryonic stem cells. Stem Cell Rev 6, 248-59.