Directed neural differentiation of induced pluripotent stem cells from non-human primates

Farnsworth, Steven L.; Qiu, Zhifang; Mishra, Anuja; Hornsby, Peter J.

doi:10.1177/1535370213482442

Cited by 12 publications

(18 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The clone that we was used in iterative process of improving differentiation (B8; Farnsworth et al, 2013) responded as expected, with increases of up to 100-fold in the tested genes. However, two other clones (88, 15) that were previously validated as iPS cells via teratoma formation (Wu et al, 2010), responded much less well (up to about 8-fold, but also failing to respond at all in some cases).…”

Section: Resultssupporting

confidence: 62%

“…We investigated how well different marmoset iPS cell clones responded to a previously developed directed differentiation protocol (Farnsworth et al, 2013). In this protocol, the cells are formed into aggregates in hanging-drop plates for 3 days and then exposed to differentiation factors (dual SMAD inhibitors plus retinoic acid) for 3 days in monolayer culture.…”

Section: Resultsmentioning

confidence: 99%

“…Subsequently we documented a rapid iterative method for developing neural cell differentiation protocols in marmoset iPS cells (Farnsworth et al, 2013). For autologous cell therapy experiments to be feasible, it must be possible to apply a differentiation protocol to iPS cell clones newly generated from donor animals without the need to customize the protocol for each cell line or for each donor.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide

Qiu

Mishra

et al. 2015

Stem Cell Research

Self Cite

View full text Add to dashboard Cite

The marmoset is an important nonhuman primate model for regenerative medicine. For experimental autologous cell therapy based on induced pluripotent (iPS) cells in the marmoset, cells must be able to undergo robust and reliable directed differentiation that will not require customization for each specific iPS cell clone. When marmoset iPS cells were aggregated in a hanging drop format for 3 days, followed by exposure to dual SMAD inhibitors and retinoic acid in monolayer culture for 3 days, we found substantial variability in the response of different iPS cell clones. However, when clones were pretreated with 0.05% – 2% dimethyl sulfoxide (DMSO) for 24 hours, all clones showed a very similar maximal response to the directed differentiation scheme. Peak responses were observed at 0.5% DMSO in two clones and at 1% DMSO in a third clone. When patterns of gene expression were examined by microarray analysis, hierarchical clustering showed very similar responses in all 3 clones when they were pretreated with optimal DMSO concentrations. The change in phenotype following exposure to DMSO and the 6 day hanging drop/monolayer treatment was confirmed by immunocytochemistry. Analysis of DNAcontent in DMSO-exposed cells indicated that it is unlikely that DMSO acts by causing cells to exit from the cell cycle. This approach should be generally valuable in the directed neural differentiation of pluripotent cells for experimental cell therapy.

show abstract

Section: Resultssupporting

confidence: 62%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide

Qiu

Mishra

et al. 2015

Stem Cell Research

Self Cite

View full text Add to dashboard Cite

show abstract

“…In fact, specific culture conditions have been developed to maintain the pluripotent stem cells (PSCs) in an extended PSC stage (EPSC) with widespread chimeric contribution to embryonic and extraembryonic lineages in vivo [ 28 ]. The derivation and tendency of the marmoset pluripotent stem cells to differentiate into neural lineages has been reported [ 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 ] using variable techniques for the neuralization based on the initial protocol developed for human embryonic stem cells [ 38 , 39 , 40 , 41 , 42 , 43 ]. In this study, we demonstrated that using the NN1 medium supplemented with growth factors, we were able to isolate NSCs that are responsive to mitogenic growth factor.…”

Section: Discussionmentioning

confidence: 99%

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

Yang

Hong

Torres

et al. 2018

IJMS

View full text Add to dashboard Cite

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.

show abstract

“…Recently, three novel marmoset ESC lines have been derived from the natural morula stage of preimplantation embryos, and one novel male ESC line has been derived from the expanded blastocyst stage (Debowski et al 2016). Marmoset iPSCs from fetal liver cells and adult fibroblasts have also been reported (Tomioka et al 2010; Farnsworth et al 2013; Qiu et al 2015; Vermilyea et al 2017). The capacity of marmoset ESC and iPSC lines to differentiate into cardiomyocytes and hematopoietic cells by induction has been demonstrated (Sasaki et al 2005; Tomioka et al 2010).…”

Section: Discussionmentioning

confidence: 99%

Haplotype-phased common marmoset embryonic stem cells for genome editing using CRISPR/Cas9

Zhou

Leung

Ward

et al. 2018

Preprint

View full text Add to dashboard Cite

Due to anatomical and physiological similarities to humans, the common marmoset (Callithrix jacchus) is an ideal organism for the study human diseases. Researchers are currently leveraging genome-editing technologies such as CRISPR/Cas9 to genetically engineer marmosets for the in vivo biomedical modeling of human neuropsychiatric and neurodegenerative diseases. The genome characterization of these cell lines greatly reinforces these transgenic efforts. It also provides the genomic contexts required for the accurate interpretation of functional genomics data. We performed haplotype-resolved whole-genome characterization for marmoset ESC line cj367 from the Wisconsin National Primate Research Center. This is the first haplotype-resolved analysis of a marmoset genome and the first whole-genome characterization of any marmoset ESC line. We identified and phased single-nucleotide variants (SNVs) and Indels across the genome. By leveraging this haplotype information, we then compiled a list of cj367 ESC allele-specific CRISPR targeting sites. Furthermore, we demonstrated successful Cas9 Endonuclease Dead (dCas9) expression and targeted localization in cj367 as well as sustained pluripotency after dCas9 transfection by teratoma assay. Lastly, we show that these ESCs can be directly induced into functional neurons in a rapid, single-step process. Our study provides a valuable set of genomic resources for primate transgenics in this post-genome era.

show abstract

Directed neural differentiation of induced pluripotent stem cells from non-human primates

Cited by 12 publications

References 29 publications

Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide

Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide

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

Haplotype-phased common marmoset embryonic stem cells for genome editing using CRISPR/Cas9

Contact Info

Product

Resources

About