Rhesus Macaque iPSC Generation and Maintenance

Yada, Ravi Chandra; Hong, So Gun; Lin, Yongshun; Winkler, Thomas; Dunbar, Cynthia E.

doi:10.1002/cpsc.25

Cited by 6 publications

(7 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We reprogrammed fibroblasts from five rhesus macaques, one baboon, and two humans to demonstrate the broad applicability of our reprogramming method. To our knowledge, most of the relatively few NHP-iPSC lines generated until now (including macaque, baboon, gorilla, chimpanzee, bonobo, and common marmoset) were typically reprogrammed using methods involving genomic integration of the reprogramming vectors [ 23 , 29 , 30 , 33 , 34 , 55 , 56 , 57 , 58 , 59 ]; or genomic integration followed by the excision of the lentiviral vector leaving a genetic tag in the host genome [ 26 , 60 ]. So far, to our knowledge, only very few studies using an approach based on non-integrating vectors were reported recently [ 16 , 21 , 24 , 27 ].…”

Section: Discussionmentioning

confidence: 99%

Non-Human Primate iPSC Generation, Cultivation, and Cardiac Differentiation under Chemically Defined Conditions

Stauske

Rodríguez-Polo

Haas

et al. 2020

Cells

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 99%

Non-Human Primate iPSC Generation, Cultivation, and Cardiac Differentiation under Chemically Defined Conditions

Stauske

Rodríguez-Polo

Haas

et al. 2020

Cells

View full text Add to dashboard Cite

show abstract

“…Rhesus CD34 + hematopoietic stem and progenitor cells or skin fibroblasts were isolated and reprogrammed as previously described 6,11,12 . RhiPSCs were cultured either on mouse embryonic fibroblast (GlobalStem) feeders or growth factor‐reduced Matrigel (BD Biosciences)‐coated plates.…”

Section: Methodsmentioning

confidence: 99%

CRISPR/Cas9-mediated introduction of the sodium/iodide symporter gene enables noninvasive in vivo tracking of induced pluripotent stem cell-derived cardiomyocytes

Ostrominski

Yada

Sato

et al. 2020

Stem Cells Translational Medicine

Self Cite

View full text Add to dashboard Cite

Techniques that enable longitudinal tracking of cell fate after myocardial delivery are imperative for optimizing the efficacy of cell-based cardiac therapies. However, these approaches have been underutilized in preclinical models and clinical trials, and there is considerable demand for site-specific strategies achieving long-term expression of reporter genes compatible with safe noninvasive imaging. In this study, the rhesus sodium/iodide symporter (NIS) gene was incorporated into rhesus macaque induced pluripotent stem cells (RhiPSCs) via CRISPR/Cas9. Cardiomyocytes derived from NIS-RhiPSCs (NIS-RhiPSC-CMs) exhibited overall similar morphological and electrophysiological characteristics compared to parental control RhiPSC-CMs at baseline and with exposure to physiological levels of sodium iodide. Mice were injected intramyocardially with 2 million NIS-RhiPSC-CMs immediately following myocardial infarction, and serial positron emission tomography/computed tomography was performed with 18 F-tetrafluoroborate to monitor transplanted cells in vivo. NIS-RhiPSC-CMs could be detected until study conclusion at 8 to 10 weeks postinjection. This NIS-based molecular imaging platform, with optimal safety and sensitivity characteristics, is primed for translation into large-animal preclinical models and clinical trials.

show abstract

“…A guide to establish optimal culture conditions for RhiPSCs can be found in UNIT 4A.11 (Yada et al, 2017). The plasmids used during transfection reactions of this protocol are adaptable to experimental requirements.…”

Section: Strategic Planningmentioning

confidence: 99%

“…Removing the puromycin resistance gene via Cre-mediated excision is important to prevent an immunogenic response if these cells are to be transplanted back into an immunocompetent donor rhesus macaque. Yada et al, 2017). For each experimental condition, prepare 6 MEF wells, and 3 MEF wells for positive and negative controls.…”

Section: Gene Targeting and Colony Isolationmentioning

confidence: 99%

See 1 more Smart Citation

CRISPR/Cas9‐Based Safe‐Harbor Gene Editing in Rhesus iPSCs

Yada

Ostrominski

Tunc

et al. 2017

CP Stem Cell Biology

Self Cite

View full text Add to dashboard Cite

NHP iPSCs provide a unique opportunity to test safety and efficacy of iPSC-derived therapies in clinically relevant NHP models. To monitor these cells in vivo, there is a need for safe and efficient labeling methods. Gene insertion into genomic safe harbors (GSHs) supports reliable transgene expression while minimizing the risk the modification poses to the host genome or target cell. Specifically, this protocol demonstrates targeting of the adeno-associated virus site 1 (AAVS1), one of the most widely used GSH loci in the human genome, with CRISPR/Cas9, allowing targeted marker or therapeutic gene insertion in rhesus macaque induced pluripotent stem cells (RhiPSCs). Furthermore, detailed instructions for screening targeted clones and a tool for assessing potential off-target nuclease activity are provided.

show abstract

Rhesus Macaque iPSC Generation and Maintenance

Cited by 6 publications

References 15 publications

Non-Human Primate iPSC Generation, Cultivation, and Cardiac Differentiation under Chemically Defined Conditions

Non-Human Primate iPSC Generation, Cultivation, and Cardiac Differentiation under Chemically Defined Conditions

CRISPR/Cas9-mediated introduction of the sodium/iodide symporter gene enables noninvasive in vivo tracking of induced pluripotent stem cell-derived cardiomyocytes

CRISPR/Cas9‐Based Safe‐Harbor Gene Editing in Rhesus iPSCs

Contact Info

Product

Resources

About