scholarly journals A New Class of Stem Cells in South Africa: Introducing Induced Pluripotent Stem cells (iPS cells)

2012 ◽  
Vol 103 (1) ◽  
pp. 16
Author(s):  
Robea Ballo ◽  
L Jacquie Greenberg ◽  
Susan H Kidson
Keyword(s):  
South Africa ◽  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
New Class ◽  
Induced Pluripotent

scholarly journals Generation of male germ cells from induced pluripotent stem cells (iPS cells): an in vitro and in vivo study

2012 ◽  
Vol 14 (4) ◽  
pp. 574-579 ◽  
Author(s):  
Yong Zhu ◽  
Hong-Liang Hu ◽  
Peng Li ◽  
Shi Yang ◽  
Wei Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
In Vivo Study ◽  
Male Germ Cells ◽  
Induced Pluripotent

scholarly journals Overcoming barriers to reprogramming and differentiation in nonhuman primate induced pluripotent stem cells

2017 ◽  
Vol 4 (2) ◽  
pp. 153-162 ◽  
Author(s):  
Jacob J. Hemmi ◽  
Anuja Mishra ◽  
Peter J. Hornsby
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Dimethyl Sulfoxide ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Cellular Reprogramming ◽  
Biological Research ◽  
Differentiation Potential ◽  
Wide Range ◽  
Induced Pluripotent

Abstract. Induced pluripotent stem cells (iPS cells) generated by cellular reprogramming from nonhuman primates (NHPs) are of great significance for regenerative medicine and for comparative biology. Autologously derived stem cells would theoretically avoid any risk of rejection due to host–donor mismatch and may bypass the need for immune suppression post-transplant. In order for these possibilities to be realized, reprogramming methodologies that were initially developed mainly for human cells must be translated to NHPs. NHP studies have typically used pluripotent cells generated from young animals and thus risk overlooking complications that may arise from generating iPS cells from donors of other ages. When reprogramming is extended to a wide range of NHP species, available donors may be middle- or old-aged. Here we have pursued these questions by generating iPS cells from donors across the life span of the common marmoset (Callithrix jacchus) and then subjecting them to a directed neural differentiation protocol. The differentiation potential of different clonal cell lines was assessed using the quantitative polymerase chain reaction. The results show that cells derived from older donors often showed less neural marker induction. These deficits were rescued by a 24 h pretreatment of the cells with 0.5 % dimethyl sulfoxide. Another NHP that plays a key role in biological research is the chimpanzee (Pan troglodytes). iPS cells generated from the chimpanzee can be of great interest in comparative in vitro studies. We investigated if similar deficits in differentiation potential might arise in chimpanzee iPS cells reprogrammed using various technologies. The results show that, while some deficits were observed in iPS cell clones generated using three different technologies, there was no clear association with the vector used. These deficits in differentiation were also prevented by a 24 h pretreatment with 0.5 % dimethyl sulfoxide.

GENERATION AND CHARACTERISATION OF INDUCED PLURIPOTENT STEM CELLS (iPSCs) FROM ADULT CANINE FIBROBLASTS

2012 ◽  
Vol 24 (1) ◽  
pp. 285
Author(s):  
Jorge A. Piedrahita ◽  
Sehwon Koh ◽  
Natasha Olby
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Es Cells ◽  
Ips Cells ◽  
Embryoid Bodies ◽  
Germ Layers ◽  
Induced Pluripotent ◽  
Derivatives Of

Pluripotent stem cells such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) can give rise to derivatives of all three germ layers and thus have great potential in regenerative medicine. In mice and humans, it has been shown that embryonic and adult fibroblasts can be reprogrammed into pluripotency by introducing four transcription factors, Oct3/4, Klf4, Sox2 and c-Myc (OKSM). In his presentation we will describe the derivation of iPS cells from adult canine fibroblast by retroviral OSKM transduction. The isolated canine iPS cells were expanded in three different iPS culture media (FGF2, LIF and FGF2 plus LIF) and only the cells cultured in FGF2 plus LIF showed strong AP activity expressed pluripotency markers, POU5F1 (OCT4), SOX2, NANOG and LIN28 as well as ES cells-specific genes (PODXL, DPPA5, FGF5, REX1 and LAMP1). In vitro differentiation by formation of embryoid bodies (EBs) and directed differentiation showed cell derivatives of all three germ layers as confirmed by expression for AFP, CXCR4 and SOX17 (endoderm), desmin (DES), vimentin (VIM), MSX1 and BMP2 (mesoderm) and glial fibrillary acidic protein (GFAP), TUJ1, NCAM and bIII-tubulin (TUBB, ectoderm). In vivo, the putative canine iPS cells formed simple teratomas that expressed markers for all three germ layers. In summary, we were able to derive induced pluripotent cells from adult somatic cells by using four transcription factors. The isolated canine iPSCs have similar characteristics to ESCs from other species, but the exact cellular mechanisms behind their unique co-dependency on both FGF and LIF is still unknown. This work was funded by a grant from the America Kennel Club to JAP.

186 Improving Success Rate of Establishment and Maintenance of Porcine Induced Pluripotent Stem Cells by Investigation of Colony Morphology

2018 ◽  
Vol 30 (1) ◽  
pp. 233
Author(s):  
P. Setthawong ◽  
P. Phakdeedindan ◽  
M. Techakumphu ◽  
T. Tharasanit
Keyword(s):  
Stem Cells ◽  
Alkaline Phosphatase ◽  
Induced Pluripotent Stem Cells ◽  
Success Rate ◽  
Cell Lines ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Fibroblast Cells ◽  
Induced Pluripotent

Induced pluripotent stem cells (iPS cells) are generated by reprogramming of somatic cells using ectopic introduction of 4 transcription factors, including OCT4, SOX2, KLF4, and c-MYC (OSKM). Fibroblast cells are the most commonly used as a primary cell source for iPS cells because they are easy to harvest and culture. However, reprogramming efficiency of porcine fibroblasts is poor (~0.1%). During reprogramming process, mixed populations of primary colonies become the major obstacle in iPS establishment. In this study, we characterised 2 different colony morphologies at colony pick-up (compact and loose iPS morphology). We hypothesised that compact colonies will proceed to long-term culture and pluripotency. The fibroblast cells were isolated from tails of piglets and transfected with retroviral vectors expressing OSKM. The primary colonies were counted on Day 7 after gene transduction. The iPS-like colonies were divided into compact (n = 10) and loose (n = 10) morphology at colony pick-up. The characteristics of iPS-like cell lines were analysed by morphology, alkaline phosphatase staining, G-banding karyotype, immunofluorescence staining (OCT4), pluripotent gene expression (RT-PCR), and embryoid body formation. A total of 1,697 iPS-like colonies (2.34%) were observed. The compact colonies contained with tightly packed cells with distinct border between iPS colony and feeder cells, while colonies with irregular shape and border were classified as loose colonies. These 2 types of iPS-like colonies expressed alkaline phosphatase and OCT4. A total 5 of 10 (50%) compact morphology cell lines could be maintained in the undifferentiated state for more than 50 passages. But only 3 of 10 (30%) loose morphology cell lines demonstrated pluripotent characteristics. G-Banding karyotype analysis revealed normal chromosome number (n = 38). All of the cell lines also expressed endogenous pluripotent genes, including OSKM and NANOG and formed three-dimensional aggregating masses. In this study, we found that the cell lines from compact morphology could be maintained for longer than those of the loose morphology. A high rate of differentiation of loose iPS colony may also indicate that this type of colony has different pluripotency signals or incomplete reprogramming compared with compact colony. In conclusion, selection of compact colony morphology at colony pick-up is simple and reflects long-term propagation of porcine iPS cell lines. This information is important for improving the success rate of establishment and maintenance of porcine iPS cells.

Induced Pluripotent Stem Cells(iPS Cells):Current Status and Future Prospect*

2009 ◽  
Vol 2009 (8) ◽  
pp. 950-960 ◽  
Author(s):  
Hong-Fen SHEN ◽  
Zhi-Fang YAO ◽  
Gao-Fang XIAO ◽  
Jun-Shuang JIA ◽  
Dong XIAO ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Future Prospect ◽  
Ips Cells ◽  
Current Status ◽  
Induced Pluripotent

Dynamic polyrotaxane-coated surface for effective differentiation of mouse induced pluripotent stem cells into cardiomyocytes

RSC Advances ◽  
2016 ◽  
Vol 6 (42) ◽  
pp. 35668-35676 ◽  
Author(s):  
Ji-Hun Seo ◽  
Mitsuhi Hirata ◽  
Sachiro Kakinoki ◽  
Tetsuji Yamaoka ◽  
Nobuhiko Yui
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Molecular Mobility ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Coated Surface ◽  
Coated Surfaces ◽  
Induced Pluripotent

Increasing molecular mobility of hydrated polyrotaxane (PRX)-coated surfaces was effective to promote the differentiation of mouse induced pluripotent stem cells (iPS cells) into cardiomyocytes.

scholarly journals Myotubes derived from human-induced pluripotent stem cells mirror in vivo insulin resistance

2016 ◽  
Vol 113 (7) ◽  
pp. 1889-1894 ◽  
Author(s):  
Salvatore Iovino ◽  
Alison M. Burkart ◽  
Laura Warren ◽  
Mary Elizabeth Patti ◽  
C. Ronald Kahn
Keyword(s):  
Insulin Resistance ◽  
Skeletal Muscle ◽  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Muscle Insulin Resistance ◽  
Induced Pluripotent

Induced pluripotent stem cells (iPS cells) represent a unique tool for the study of the pathophysiology of human disease, because these cells can be differentiated into multiple cell types in vitro and used to generate patient- and tissue-specific disease models. Given the critical role for skeletal muscle insulin resistance in whole-body glucose metabolism and type 2 diabetes, we have created a novel cellular model of human muscle insulin resistance by differentiating iPS cells from individuals with mutations in the insulin receptor (IR-Mut) into functional myotubes and characterizing their response to insulin in comparison with controls. Morphologically, IR-Mut cells differentiated normally, but had delayed expression of some muscle differentiation-related genes. Most importantly, whereas control iPS-derived myotubes exhibited in vitro responses similar to primary differentiated human myoblasts, IR-Mut myotubes demonstrated severe impairment in insulin signaling and insulin-stimulated 2-deoxyglucose uptake and glycogen synthesis. Transcriptional regulation was also perturbed in IR-Mut myotubes with reduced insulin-stimulated expression of metabolic and early growth response genes. Thus, iPS-derived myotubes from individuals with genetically determined insulin resistance demonstrate many of the defects observed in vivo in insulin-resistant skeletal muscle and provide a new model to analyze the molecular impact of muscle insulin resistance.

scholarly journals Patient-Specific Induced Pluripotent Stem Cells for SOD1-Associated Amyotrophic Lateral Sclerosis Pathogenesis Studies

Acta Naturae ◽  
2014 ◽  
Vol 6 (1) ◽  
pp. 54-60 ◽  
Author(s):  
I. V. Chestkov ◽  
E. A. Vasilieva ◽  
S. N. Illarioshkin ◽  
M. A. Lagarkova ◽  
S. L. Kiselev
Keyword(s):  
Stem Cells ◽  
Amyotrophic Lateral Sclerosis ◽  
Induced Pluripotent Stem Cells ◽  
Motor Neurons ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
The Body ◽  
Patient Specific ◽  
Induced Pluripotent ◽  
Lateral Sclerosis

The genetic reprogramming technology allows one to generate pluripotent stem cells for individual patients. These cells, called induced pluripotent stem cells (iPSCs), can be an unlimited source of specialized cell types for the body. Thus, autologous somatic cell replacement therapy becomes possible, as well as the generation of in vitro cell models for studying the mechanisms of disease pathogenesis and drug discovery. Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disorder that leads to a loss of upper and lower motor neurons. About 10% of cases are genetically inherited, and the most common familial form of ALS is associated with mutations in the SOD1 gene. We used the reprogramming technology to generate induced pluripotent stem cells with patients with familial ALS. Patient-specific iPS cells were obtained by both integration and transgene-free delivery methods of reprogramming transcription factors. These iPS cells have the properties of pluripotent cells and are capable of direct differentiation into motor neurons.

326 ASSESSMENT OF PORCINE-INDUCED PLURIPOTENT STEM CELLS BY IN VIVO ASSAYS

2015 ◽  
Vol 27 (1) ◽  
pp. 252
Author(s):  
J. Secher ◽  
K. Freude ◽  
S. Petkov ◽  
A. Ceylan ◽  
M. Schmidt ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Differential Staining ◽  
Good Survival ◽  
Vitro Assay ◽  
Induced Pluripotent

Porcine-induced pluripotent stem cells (piPSC) have been established since 2009, but only 1 report demonstrated contribution to germline chimeras. One well-established in vivo pluripotency assay is the teratoma assay, which has recently been questioned due to the lack of standardized guidelines. In the present study we have characterised GFP-tagged in vitro and in vivo tetracycline-inducible piPSC [porcine MYC, SOX2, KLF4 (pOMSK)] and their capacity to form teratomas. We injected 1.5 million cells in 250 µL of PBS subcutaneous into NOD/SCID mice and followed them up to 6 weeks. The teratomas were analysed by immunohistochemistry for the 3 germlayer markers β3 tubulin, α fetoprotein, and smooth muscle actin. We not only found our teratomas positive for these markers, but also co-positive for GFP, clearly showing that the teratoma was made from porcine cells, which was not sufficiently proven in former studies. Our H&E staining revealed the following structures: cuboidal ephitelium, thyroid-like structure, renal corpuscle, and steroid producing cells. We continued to test the capacity of our venus iPS cells to contribute to in vitro chimeras. To achieve this we used a micromanipulator to inject 15 cells into Day 5 parthenotes, and subsequently cultured them in PZM3 with 10% FCS, cultured with or without doxycycline. These in vitro chimeras were followed until Day 7 in Nikons Biostation IM and used for differential staining. In all groups we saw good survival, hatching, and maintenance of GFP, indicating integration of these cells in our in vitro assay. We only found differences between survivals of the cell lines in the group cultured with doxycycline. Finally, in order to assess if the naïve type venus iPS cells could possibly be a truly naïve piPSC, we tested their capacity to form in vivo germline chimeras. This was tested by injecting 15 piPSC into Day 4 to 5 in vivo embryos. The injected embryos were transferred into 5 surrogate mothers, 3 of them were fed doxycycline before the transfer and 5 days after, and the last 2 recipient sows were not fed doxycycline. The pregnancies were terminated at Day 32 and the embryos were examined for fluorescence and the GFP transgene by PCR. In summary, it appears that both naïve type and primed type venus iPS cells are still strongly dependent on the pOMSK transgene expression, and the ultimate proof for pluripotency, the chimera production, seems to be not achievable under the condition we have chosen.

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