Bioluminescent imaging demonstrates that transplanted human embryonic stem cell-derived CD34(+) cells preferentially develop into endothelial cells.

Tian X1, Hexum MK, Penchev VR, Taylor RJ, Shultz LD, Kaufman DS.
Source: Stem Cells
Publication Date: (2009)
Issue: 27(11): 2675(85)
Research Area:
Stem Cells
Basic Research
Cells used in publication:
Embryonic Stem Cell (ES), human
Species: human
Tissue Origin: embryo
Nucleofector™ I/II/2b
Nucleofection of hES cells. Suboptimal kit selection, they used Cell Line Kit V instead of human stem cell kit. Undifferentiated hESCs were maintained on Matrigel-coated plates for at least 1 passage before nucleofection. Small clumps of hESCs were harvested and nucleofected with 5–10 µg of transposon DNA in combination with 2.5–5 µg of transposase DNA in 100 µl of nucleofector solution V using program setting B16 as previously described. Nucleofected hESCs were immediately transferred onto irradiated mouse embryonic fibroblasts (MEF) with pre-warmed hESC growth medium as previously described.
Human embryonic stem cells (hESCs) provide an important resource for novel regenerative medicine therapies and have been used to derive diverse cell populations, including hematopoietic and endothelial cells. However, it remains a challenge to achieve significant engraftment of hESC-derived blood cells when transplanted into animal models. To better understand mechanisms that enhance or limit the in vivo developmental potential of hESC-derived cells, we utilized hESCs that express firefly luciferase (luc) to allow noninvasive, real-time bioluminescent imaging of hESC-derived CD34(+) cells transplanted into the liver of neonatal immunodeficient mice. Serial imaging demonstrated stable engraftment and expansion of the luc(+) hESC-derived cells in vivo over several months. While we found that these hESC-derived CD34(+) cells have bipotential ability to generate both hematopoietic and endothelial lineages in vitro, these studies demonstrate preferential differentiation into endothelial cells in vivo, with only low levels of hematopoietic cell engraftment. Therefore, these studies reveal key differences in the developmental potential of hESC-derived cells using in vitro and in vivo analyses. Although transplanted hESC-derived CD34(+) cells are well-suited for revascularization therapies, additional measures are needed to provide higher levels of long-term hematopoietic engraftment.