Journal: bioRxiv

Article Title: Cross-species blastocyst chimerism between nonhuman primates using iPSCs

doi: 10.1101/635250

Figure Lengend Snippet: A) Colony morphology of pig-tailed macaque iPSC in NHP-E8X media on MEF at 4X and 10X resolution (left) and the colony morphology of pig-tailed macaque iPSCs in E8 media on MEF at 4X and 10X resolution (right). B) Co-culture of human-human iPSC (top-left) and human-chimp iPSC (top-right). Human and chimpanzee iPSCs mix and integrate homogenously without forming segregated colonies. Co-culture of mouse-mouse iPSCs form mixed and homogenously integrated colonies (bottom-left). Human and mouse iPSCs form separate and segregated colonies (bottom-right). C) Cardiomyocyte layers differentiated from the co-culture mixture of chimpanzee and human iPSCs. The tdTomato-fluorescent labeled beating chimpanzee iPSC (red, left) and the human GFP-fluorescent labeled beating cardiomyocyte (green, middle) and the overlay image of the mixed cardiomyocytes from the mixture of chimpanzee and human iPSC (Supplementary video 1). Electrocardiogram of cardiomyocyte derived from the mixture of human and chimpanzee iPSCs indicating contraction and relaxation start, peak, and end (right). D) Hematoxylin and eosin (H&E)-stained sections of chimpanzee teratoma showing all three embryonic germ layers, derived from chimpanzee iPSCs with and without Bcl2 transduction. Ectoderm (a’, b’) is characterized by the presence of neural ectoderm arranged in rosette-like patterns (black arrows, a’ and b’), that resemble embryonic neural tubes (white arrows, a’). Endoderm (a’’, b’’) is characterized by the presence of glandular and/or secretory epithelium (black asterisks). Mesoderm (a’’’, b’’’) is characterized by the presence of elongate, mesenchymal cells (black arrows, a’’’) and cells embedded in a pale blue gray hyaline matrix resembling articular cartilage (black asterisk, b’’’). Magnification: 40x. Scale bars: 20μm.

Article Snippet: On the day after transfection (day 1), all the cells transfected with Sendai virus were transferred into a 6-well plate (30,000 cells per well) each well containing ~150,000 mouse embryonic fibroblast (MEF) feeder cells (Stemcell Technologies Inc. Vancouver, Canada).

Techniques: Co-Culture Assay, Labeling, Derivative Assay, Staining, Transduction