Cardiomyocytes

 


A fibroblast-derived, integration-free iPSC line generated using episomal plasmids was differentiated to cardiomyocytes. Cells were exposed to RPMI/B27 (insulin-free) containing 5µM CHIR99021 on day 1, and 12µM IWP4 on days 3 and 4 (cells were fed with RPMI/B27 without small molecules on days 2, 5, and 6). On day 7, media was changed to RPMI/B27 (plus insulin) and was changed every 3 days until beating sheets of cardiomyocytes were evident. Immunofluorescence analysis of the cardiomyocyte marker cardiac troponin type 2 (Tnnt2) exhibiting typical striated staining pattern and video of spontaneously contracting cardiomyocytes are shown.

Lian et al., Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A, 2012.

Macrophages


 
A PBMC-derived iPSC line was generated using a lentiviral reprogramming vector. iPSCs were co-cultured with OP9 cells for 9 days and GM-CSF/M-CSF for 2 days followed by isolation of CD235a-/CD41a-/CD45+ cells. These cells were terminally differentiated into macrophages by exposure to GM-CSF and M-CSF for an additional 7 days. Cytospin and Wright-Geimsa staining was performed to demonstrate generation of cells with macrophage morphology. Macrophage identity of generated cells was confirmed by immunophenotyping and functional analysis.

Choi, K. D., et al., Generation of mature human myelomonocytic cells through expansion and differentiation of pluripotent stem cell-derived lin-CD34+CD43+CD45+ progenitors. J Clin Invest 119(9): 2818-2829, 2009.