Standards for Deriving Nonhuman Primate-Induced Pluripotent Stem Cells, Neural Stem Cells and Dopaminergic Lineage

Abstract

Humans and nonhuman primates (NHP) are similar in behavior and in physiology, specifically the structure, function, and complexity of the immune system. Thus, NHP models are desirable for pathophysiology and pharmacology/toxicology studies. Furthermore, NHP-derived induced pluripotent stem cells (iPSCs) may enable transformative developmental, translational, or evolutionary studies in a field of inquiry currently hampered by the limited availability of research specimens. NHP-iPSCs may address specific questions that can be studied back and forth between in vitro cellular assays and in vivo experimentations, an investigational process that in most cases cannot be performed on humans because of safety and ethical issues. The use of NHP model systems and cell specific in vitro models is evolving with iPSC-based three-dimensional (3D) cell culture systems and organoids, which may offer reliable in vitro models and reduce the number of animals used in experimental research. IPSCs have the potential to give rise to defined cell types of any organ of the body. However, standards for deriving defined and validated NHP iPSCs are missing. Standards for deriving high-quality iPSC cell lines promote rigorous and replicable scientific research and likewise, validated cell lines reduce variability and discrepancies in results between laboratories. We have derived and validated NHP iPSC lines by confirming their pluripotency and propensity to differentiate into all three germ layers (ectoderm, mesoderm, and endoderm) according to standards and measurable limits for a set of marker genes. The iPSC lines were characterized for their potential to generate neural stem cells and to differentiate into dopaminergic neurons. These iPSC lines are available to the scientific community. NHP-iPSCs fulfill a unique niche in comparative genomics to understand gene regulatory principles underlying emergence of human traits, in infectious disease pathogenesis, in vaccine development, and in immunological barriers in regenerative medicine.

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

“…It is generally recognized that the reprogramming of NHP cells is less efficient in comparison to the well-established human reprogramming techniques [ 31 , 32 ], although this has not been systematically quantified so far. For instance, it has been demonstrated that marmoset monkey iPSC generation required five or even six reprogramming factors [ 33 , 34 , 35 ], and marmoset cell reprogramming with the four Yamanaka factors was successful only after around 100 days [ 36 ]. In agreement with these observations, also low efficiency of macaque iPSC line generation has been recognized [ 23 ].…”

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

“…As study models, NH-primates are highly attractive due to longevity, behavioral, anatomical, genetic, physiological, and immunological similarities with humans [5,[7][8][9][10]. Over the past decades, NH-primates have been used on studies and research to prevent or cure human conditions, through the development of vaccines and drugs or treatment for cancer, diabetes, obesity, Parkinson's and other neurodegenerative, respiratory and cardiovascular diseases [4,5,11,12], as well as methods to prevent mother-fetus transmission of diseases such as HIV [13], amongst other conditions and illnesses.…”

“…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.…”

“…Huntington's disease transgenic animals iPSCs have been used for generating neural progenitor cells that may be addressed for drug screening [48] pathogenesis modeling [40] and epigenetic and transcriptional profile analyses [49]. iPSCs and iPSCs-derived neural stem cells have also been generated from other NH-primate species aiming to develop regenerative therapy methods and modeling other neurological conditions, such as Alzheimer's and Parkinson's Disease [10,36,[50][51][52].…”

Highly efficient induction of primate iPS cells by combining RNA transfection and chemical compounds

“…iPS cells have been generated from fetus liver‐, newborn skin‐ and adult bone marrow‐derived cells by introducing reprogramming factors using retrovirus or lentivirus (Tomioka et al, ; Wiedemann et al, ; Wu, Zhang, Mishra, Tardif, & Hornsby, ). More recently, iPS cells were generated without altering the genomic sequence using an excisable transposon vector and episomal vector from postnatal and adult skin cells, respectively (Debowski et al, ; Yang et al, ). In humans, RNA‐based methods are increasingly applied in the integration‐free generation of iPS cells, as they can lower the risk of introducing mutation (Anokye‐Danso et al, ; Mandal & Rossi, ; Poleganov et al, ; Rohani et al, ; Warren et al, ; Yoshioka et al, ).…”