Establishment of a translational endothelial cell model using directed differentiation of induced pluripotent stem cells from Cynomolgus monkey

Abstract

Due to their broad differentiation potential, pluripotent stem cells (PSCs) offer a promising approach for generating relevant cellular models for various applications. While human PSC-based cellular models are already advanced, similar systems for non-human primates (NHPs) are still lacking. However, as NHPs are the most appropriate animals for evaluating the safety of many novel pharmaceuticals, the availability of in vitro systems would be extremely useful to bridge the gap between cellular and animal models. Here, we present a NHP in vitro endothelial cell system using induced pluripotent stem cells (IPSCs) from Cynomolgus monkey (Macaca fascicularis). Based on an adapted protocol for human IPSCs, we directly differentiated macaque IPSCs into endothelial cells under chemically defined conditions. The resulting endothelial cells can be enriched using immuno-magnetic cell sorting and display endothelial marker expression and function. RNA sequencing revealed that the differentiation process closely resembled vasculogenesis. Moreover, we showed that endothelial cells derived from macaque and human IPSCs are highly similar with respect to gene expression patterns and key endothelial functions, such as inflammatory responses. These data demonstrate the power of IPSC differentiation technology to generate defined cell types for use as translational in vitro models to compare cell type-specific responses across species.

Induced pluripotent stem cells throughout the animal kingdom: Availability and applications

“…Further iPSCs have been generated from cynomolgus monkeys[156,157]. Cynomolgus monkeys are commonly used in biomedical research and the described iPSCs have been derived using both retroviral approaches[156] and non-integrative Sendai virus[157] approaches.…”

“…Further iPSCs have been generated from cynomolgus monkeys[156,157]. Cynomolgus monkeys are commonly used in biomedical research and the described iPSCs have been derived using both retroviral approaches[156] and non-integrative Sendai virus[157] approaches. Whilst in the retroviral approach pluripotency was confirmed via teratoma assays, the Sendai virus reprogrammed cells were not subjected to pluripotency assays and were directly differentiated into the cell type of interest (Supplemental material 8).…”

Angelman syndrome patient neuron screen identifies a potent and selective clinical ASO targeting UBE3A-ATS with long lasting effect in cynomolgus monkey

“…Angelman syndrome (clinicaltrials.gov; NCT04428281). NPCs derived from cynomolgus macaque (Cips_BC671A_NSC line generated at Roche) were differentiated in NPC differentiation medium (42). Differentiating neurons were incubated at indicated concentration from day 15 with ASO.…”

Induced Pluripotent Stem Cells from Animal Models: Applications on Translational Research

“…Amongst NH-primates, they include but are not limited to the rhesus [27]; drill [28]; cynomolgus monkey [29,30]; marmoset [31]; baboon [32]; orangutans [33]; Japanese macaque [34]. These cells were mainly generated from fibroblasts and integrative methods, but more recently, they were produced through non-integrative methods, such as Sendai-virus and episomal vectors [10,[35][36][37]. NH-primate-derived iPSCs have been used in research related to or as models for neurological [38][39][40][41], cardiac [36,42,43], reproductive [44], hematopoietic conditions [37,45], transplantation and grafting [30,46] and others.…”