Human hemangioblasts exist only during the early embryonic developmental stage thereby limiting the adult cellular source from which to obtain such cells for study. To overcome this, hemangioblast studies have focused on utilizing human embryonic stem cell (hESC) derivatives but current methods are cell-line dependent. Single cell dissociation of a hESC colony quickly led to cell death in most hESC lines due to enzyme treatment which, in turn, reduced induction potential and hemangioblast differentiation efficiency. Therefore, we sought to effectively improve the process of cell dissociation that is adaptable to various hESC lines and increase the initial induction potential of embryoid body (hEB). As a result, we determined an effective cell dissociation method through a comparison study involving various reagents which demonstrated successful dissociation regardless of cell line and enhanced hemangioblast differentiation efficiency.
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This study was supported by grant (2011–0019487) from the Bio & Medical Technology Development Program of the National Research Foundation (NRF), and supported by a grant (PJ.00995602) (PJ.00933303) from the Next-Generation BioGreen 21 Program of Rural Development Administration funded by the Korean government.
Supplementary Fig. 1—(A) Captured live image of the cell dissociation process with DPBS which begins at the outer edge of the colony and (B) suriving single cells.
Supplementary Fig. 2—Hemangioblast derivation from CHA15-hESCs using DPBS method. (A) Colonized CHA15-hESCs (B) EB formation (C) derived hemangioblasts and (D) commitment to blood-lineage.
Supplementary Live Image 1—Live image exhibiting the DPBS dissociation process (×4) and captured images at 0, 5, and 10 min.
Sung-Hwan Moon, Hoon-Taek Lee and Hyung-Min Chung authors have contributed equally to this paper as corresponding authors.
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Gil, C., Lee, J., Seo, J. et al. Well-defined differentiation of hesc-derived hemangioblasts by embryoid body formation without enzymatic treatment. Biotechnol Lett 37, 1315–1322 (2015). https://doi.org/10.1007/s10529-015-1786-4
- Cell dissociation
- Embryonic development
- Human pluripotent stem cells