Abstract:IntroductionZellweger spectrum disorder (PBD-ZSD) is a disease continuum caused by mutations in a subset of PEX genes required for normal peroxisome assembly and function. They highlight the importance of peroxisomes in the development and functions of the central nervous system, liver, and other organs. To date, the underlying bases for the cell-type specificity of disease are not fully elucidated.MethodsPrimary skin fibroblasts from seven PBD-ZSD patients with biallelic PEX1, PEX10, PEX12, or PEX26 mutations… Show more
“…We provide alignments of human probe sequences corresponding to these pluripotency genes and orangutan genomic sequences in Additional file 2 . Overall, the orangutan gene profiling results were consistent with that of human iPSCs produced in our laboratory at approximately the same time [ 37 , 38 ]. Fig.…”
Section: Resultssupporting
confidence: 74%
“…Figure 2 depicts the histology of one of these teratomas, which demonstrated the presence of tissues representative of all three germ layers. The results of in vitro and in vivo differentiation assays conducted on orangutan iPSCs were indistinguishable from those of human iPSCs produced in our laboratory at approximately the same time [ 37 , 38 ]. Fig.…”
Section: Resultsmentioning
confidence: 92%
“…Primary skin fibroblast cultures previously derived from punch biopsies of the upper limbs of two Sumatran orangutans (KB10973, 29 year old male; KB10460, 43 year old female) [ 33 ] were obtained from the Zoological Society of San Diego. They were transduced with retroviruses designed to express the human OCT4 , SOX2 , KLF4 and c - MYC genes, as previously described [ 37 , 38 ] (“ Methods ” Section). We observed iPSC-like colonies by 2 weeks and clonally expanded TRA-1–60 positive colonies by 3 weeks, consistent with prior reports of reprogramming skin fibroblasts from healthy human donors [ 37 , 39 ].…”
Section: Resultsmentioning
confidence: 99%
“…1 ). No differences were observed in orangutan iPSC with respect the colony morphology, passage characteristics, or molecular characteristics relative to other human iPSCs that were produced in our laboratory at approximately the same time [ 37 , 38 ]. Fig.…”
BackgroundOrangutans are an endangered species whose natural habitats are restricted to the Southeast Asian islands of Borneo and Sumatra. Along with the African great apes, orangutans are among the closest living relatives to humans. For potential species conservation and functional genomics studies, we derived induced pluripotent stem cells (iPSCs) from cryopreserved somatic cells obtained from captive orangutans.ResultsPrimary skin fibroblasts from two Sumatran orangutans were transduced with retroviral vectors expressing the human OCT4, SOX2, KLF4, and c-MYC factors. Candidate orangutan iPSCs were characterized by global gene expression and DNA copy number analysis. All were consistent with pluripotency and provided no evidence of large genomic insertions or deletions. In addition, orangutan iPSCs were capable of producing cells derived from all three germ layers in vitro through embryoid body differentiation assays and in vivo through teratoma formation in immune-compromised mice.ConclusionsWe demonstrate that orangutan skin fibroblasts are capable of being reprogrammed into iPSCs with hallmark molecular signatures and differentiation potential. We suggest that reprogramming orangutan somatic cells in genome resource banks could provide new opportunities for advancing assisted reproductive technologies relevant for species conservation efforts. Furthermore, orangutan iPSCs could have applications for investigating the phenotypic relevance of genomic changes that occurred in the human, African great ape, and/or orangutan lineages. This provides opportunities for orangutan cell culture models that would otherwise be impossible to develop from living donors due to the invasive nature of the procedures required for obtaining primary cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1567-0) contains supplementary material, which is available to authorized users.
“…We provide alignments of human probe sequences corresponding to these pluripotency genes and orangutan genomic sequences in Additional file 2 . Overall, the orangutan gene profiling results were consistent with that of human iPSCs produced in our laboratory at approximately the same time [ 37 , 38 ]. Fig.…”
Section: Resultssupporting
confidence: 74%
“…Figure 2 depicts the histology of one of these teratomas, which demonstrated the presence of tissues representative of all three germ layers. The results of in vitro and in vivo differentiation assays conducted on orangutan iPSCs were indistinguishable from those of human iPSCs produced in our laboratory at approximately the same time [ 37 , 38 ]. Fig.…”
Section: Resultsmentioning
confidence: 92%
“…Primary skin fibroblast cultures previously derived from punch biopsies of the upper limbs of two Sumatran orangutans (KB10973, 29 year old male; KB10460, 43 year old female) [ 33 ] were obtained from the Zoological Society of San Diego. They were transduced with retroviruses designed to express the human OCT4 , SOX2 , KLF4 and c - MYC genes, as previously described [ 37 , 38 ] (“ Methods ” Section). We observed iPSC-like colonies by 2 weeks and clonally expanded TRA-1–60 positive colonies by 3 weeks, consistent with prior reports of reprogramming skin fibroblasts from healthy human donors [ 37 , 39 ].…”
Section: Resultsmentioning
confidence: 99%
“…1 ). No differences were observed in orangutan iPSC with respect the colony morphology, passage characteristics, or molecular characteristics relative to other human iPSCs that were produced in our laboratory at approximately the same time [ 37 , 38 ]. Fig.…”
BackgroundOrangutans are an endangered species whose natural habitats are restricted to the Southeast Asian islands of Borneo and Sumatra. Along with the African great apes, orangutans are among the closest living relatives to humans. For potential species conservation and functional genomics studies, we derived induced pluripotent stem cells (iPSCs) from cryopreserved somatic cells obtained from captive orangutans.ResultsPrimary skin fibroblasts from two Sumatran orangutans were transduced with retroviral vectors expressing the human OCT4, SOX2, KLF4, and c-MYC factors. Candidate orangutan iPSCs were characterized by global gene expression and DNA copy number analysis. All were consistent with pluripotency and provided no evidence of large genomic insertions or deletions. In addition, orangutan iPSCs were capable of producing cells derived from all three germ layers in vitro through embryoid body differentiation assays and in vivo through teratoma formation in immune-compromised mice.ConclusionsWe demonstrate that orangutan skin fibroblasts are capable of being reprogrammed into iPSCs with hallmark molecular signatures and differentiation potential. We suggest that reprogramming orangutan somatic cells in genome resource banks could provide new opportunities for advancing assisted reproductive technologies relevant for species conservation efforts. Furthermore, orangutan iPSCs could have applications for investigating the phenotypic relevance of genomic changes that occurred in the human, African great ape, and/or orangutan lineages. This provides opportunities for orangutan cell culture models that would otherwise be impossible to develop from living donors due to the invasive nature of the procedures required for obtaining primary cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1567-0) contains supplementary material, which is available to authorized users.
“…Cultured patient cells, including skin fibroblasts, have provided invaluable for screening and testing drug therapies in vitro [78]. Most recently, PBD-ZSD patient-derived skin fibroblasts have been reprogrammed into induced pluripotent stem cells (iPSCs) that were differentiated into neural and hepatic cell models of disease that could be used in drug screening and testing efforts [79]. There are several genetically engineered mouse models of PEX gene defects [80], including a model of the common PEX1 p.G843D mutation [81].…”
Peroxisome biogenesis disorders in the Zellweger spectrum (PBD-ZSD) are a heterogeneous group of genetic disorders caused by mutations in PEX genes responsible for normal peroxisome assembly and functions. As a result of impaired peroxisomal activities, individuals with PBD-ZSD can manifest a complex spectrum of clinical phenotypes that typically result in shortened life spans. The extreme variability in disease manifestation ranging from onset of profound neurologic symptoms in newborns to progressive degenerative disease in adults presents practical challenges in disease diagnosis and medical management. Recent advances in biochemical methods for newborn screening and genetic testing have provided unprecedented opportunities for identifying patients at the earliest possible time and defining the molecular bases for their diseases. Here, we provide an overview of current clinical approaches for the diagnosis of PBD-ZSD and provide broad guidelines for the treatment of disease in its wide variety of forms. Although we anticipate future progress in the development of more effective targeted interventions, the current guidelines are meant to provide a starting point for the management of these complex conditions in the context of personalized health care.
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