Abstract
Non-human primates (NHP) are important surrogate models for late preclinical development of advanced therapy medicinal products (ATMPs), including induced pluripotent stem cell (iPSC)-based therapies, which are also under development for heart failure repair. For effective heart repair by remuscularization, large numbers of cardiomyocytes are required, which can be obtained by efficient differentiation of iPSCs. However, NHP-iPSC generation and long-term culture in an undifferentiated state under feeder cell-free conditions turned out to be problematic. Here we describe the reproducible development of rhesus macaque (Macaca mulatta) iPSC lines. Postnatal rhesus skin fibroblasts were reprogrammed under chemically defined conditions using non-integrating vectors. The robustness of the protocol was confirmed using another NHP species, the olive baboon (Papio anubis). Feeder-free maintenance of NHP-iPSCs was essentially dependent on concurrent Wnt-activation by GSK-inhibition (Gi) and Wnt-inhibition (Wi). Generated NHP-iPSCs were successfully differentiated into cardiomyocytes using a combined growth factor/GiWi protocol. The capacity of the iPSC-derived cardiomyocytes to self-organize into contractile engineered heart muscle (EHM) was demonstrated. Collectively, this study establishes a reproducible protocol for the robust generation and culture of NHP-iPSCs, which are useful for preclinical testing of strategies for cell replacement therapies in NHP.
Highlights
The value of non-human primates (NHP) as animal models in translational research is increasinglyThe value of non-human primates (NHP) as animal models in translational research is recognized [1]
To quantify and compare reprogramming efficiencies, we investigated reprogramming in primary fibroblast cultures obtained from three adult rhesus macaques and two human adults
Primary colonies were counted after staining for alkaline phosphatase (AP) at days 20, 30, and 40 after nucleofection with the reprogramming factors
Summary
The value of non-human primates (NHP) as animal models in translational research is increasingly. The value of non-human primates (NHP) as animal models in translational research is recognized [1]. NHP offer advantages compared to other species because of their close phylogenetic increasingly recognized [1]. NHP offer advantages compared to other species because of their close relationship to humans [2,3,4] (Figure 1). Phylogenetic relationship to humans [2,3,4]( Figure 1). Non-human primates primates in in basic basic and and translational translational research. Phylogenetic tree tree of of mammals mammals used as model organisms in research (after [5] ),
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