Human Induced Pluripotent Stem Cells: The Past, Present, and Future

Biotechnology Journal, 2012

Various methods of iPS cell generation have been developed to improve the quality of iPS cells, the efficiency of reprogramming and the feasibility and safety of using iPS cells for clinical applica

Stem Cell Research, 2016

II is a healthy feeder-free and characterized human induced pluripotent stem (iPS) cell line. Cultured under xeno-free and defined conditions. The line is generated from healthy human fibroblasts with nonintegrating Sendai virus vectors encoding the four Yamanaka factors, OCT4, SOX2, KLF4 and cMYC. The generated iPS cells are free from reprogramming vectors and their purity, karyotypic stability and pluripotent capacity is confirmed.

2014

Undoubtedly, the focus of the field of stem cell research is predominantly aimed at the artificial reprogramming of human somatic cells for the production of induced pluripotent stem (iPS) cells. This relatively new technology may circumvent the ethical issues of using human embryonic stem (hES) cells for the potential applications in cell replacement therapy. Besides such ethical issues, iPS cell technology offers the advantage of obtaining patient-derived tissues and/or cells, which may be utilized for autologous transplantation and tissue regeneration, investigation of a variety of human illnesses and for the screening of new drugs. The field of stem cell research has placed a major emphasis in understanding the genetic and epigenetic codes for pluripotency, in order to control and optimize autologous transplantation techniques and avoid teratoma formation.

International Journal of Stem cell Research & Therapy, 2016

A decade has passed since Shinya Yamanaka published his landmarking publication on how to produce pluripotent stemcell-like cells and the term induced pluripotent stem cells, iPS cells, was coined. This past decade has been a decade devoted to pluripotent cells, scrutinizing molecular mechanisms in cell identity, and optimizing derivation methods, culture conditions and characterization methods to xenofree and chemically defined clinical-grade pluripotent stem cells. The early promises of autologous cell therapies are now replaced by creation of highly selected donor cell banks matched to provide cells for the majority of a target population. Several of the initial safety concerns with iPS cells have been addressed with the use of non-integrating derivation methods and chemically defined and xenofree culture conditions, but some remain and will not be fully resolved until conclusions from in vivo experiments in larger animal models can be made. Published studies on safety and proof of concept performed in nonhuman primates are few but show promising results for spinal cord injury and Parkinson's disease for example. But questions remain; on how to provide functional and long-term integrating grafts and whether these can fulfill the promises of recovery and potential cure?

Science China Life Sciences, 2011

Induced pluripotent stem (iPS) cell technology demonstrates that somatic cells can be reprogrammed to a pluripotent state by over-expressing four reprogramming factors. This technology has created an interest in deriving iPS cells from domesticated animals such as pigs, sheep and cattle. Moloney murine leukemia retrovirus vectors have been widely used to generate and study mouse iPS cells. However, this retrovirus system infects only mouse and rat cells, which limits its use in establishing iPS cells from other mammals. In our study, we demonstrate a novel retrovirus strategy to efficiently generate porcine iPS cells from embryonic fibroblasts. We transfected four human reprogramming factors (Oct4, Sox2, Klf4 and Myc) into fibroblasts in one step by using a VSV-G envelope-coated pantropic retrovirus that was easily packaged by GP2-293 cells. We established six embryonic stem (ES)-like cell lines in human ES cell medium supplemented with bFGF. Colonies showed a similar morphology to human ES cells with a high nuclei-cytoplasm ratio and phase-bright flat colonies. Porcine iPS cells could form embryoid bodies in vitro and differentiate into the three germ layers in vivo by forming teratomas in immunodeficient mice. induced pluripotent stem cells, Moloney murine leukemia retrovirus vectors, embryoid body, teratoma

Nature Protocols, 2010

Induced Pluripotent Stem Cells, 2012

Philosophical Transactions of the Royal Society B: Biological Sciences, 2011

Somatic cells have been reprogrammed into pluripotent stem cells by introducing a combination of several transcription factors, such as Oct3/4, Sox2, Klf4, and c-Myc.

Journal of Assisted Reproduction and Genetics, 2011

Direct reprogramming of somatic cells into induced pluripotent stem (iPS) cells has emerged as an invaluable method for generating patient-specific stem cells of any lineage without the use of embryonic materials. Following the first reported generation of iPS cells from murine fibroblasts using retroviral transduction of a defined set of transcription factors, various new strategies have been developed to improve and refine the reprogramming technology. Recent developments provide optimism that the generation of safe iPS cells without any genomic modification could be derived in the near future for the use in clinical settings. This review summarizes current and evolving strategies in the generation of iPS cells, including types of somatic cells for reprogramming, variations of reprogramming genes, reprogramming methods, and how the advancement iPS cells technology can lead to the future success of reproductive medicine.

Embryonic Stem Cells - Differentiation and Pluripotent Alternatives, 2011

Regenerative Medicine, 2010

Stem Cell Reviews and Reports, 2012

Cellular reprogramming consists of the conversion of differentiated cells into pluripotent cells; the so-called induced Pluripotent Stem Cells. iPSC are amenable to in vitro manipulation and, in theory, direct production of any differentiated cell type. Furthermore, iPSC can be obtained from sick individuals and subsequently used for disease modeling, drug discovery and regenerative treatments. iPSC production was first achieved by transducing, with the use of retroviral vectors, four specific transcription factors: Oct4, Klf4, Sox2 and c-Myc (OKSM), into primary cells in culture Takahashi and Yamanaka, (Cell 126(4):663-676, 2006). Many alternative protocols have since been proposed: repeated transfections of expression plasmids containing the four pluripotencyassociated genes Okita et al.

Journal of stem cells & regenerative medicine, 2013

Cell therapy is one of the principal and most promising research areas of regenerative medicine. Nevertheless different ethical problems related with the use of embryonic stem cells arise from this technology. Thus the search for a feasible way of stem cells generation is in current investigation. Induced pluripotent stem (iPS) cell is one of the most encouraging emerging technologies that represent a solution to this problem. Different methods that change cell protein and gene profiles, as well as its morphology and function, driving the cell to the stem cell state have been developed. These include nuclear transfer, the usage of cell extracts and synthetic molecules, the forced expression of defined genes and cytoplasmatic level modifications. Even though favorable results have been achieved, there are still issues that require special attention. Advances with non-integration methods are now available but dedifferentiation efficiency is yet an area of opportunity. The main objecti...

Cell Stem Cell, 2009

Nature Methods, 2011

Table 8). Theoretically, iPSCs established from these two individuals match ~20% of all the combinations of 2,117 haplotypes in Japanese population. Indeed, pla-iPSC lines derived from lines DP74 and DP94 match 32 of 107 donors 11 at the three HLA loci (HLA-A, HLA-B and HLA-DR) with the two-digit specification (Supplementary Table 7). Others previously estimated that iPSC lines with 50 unique HLA homozygous haplotypes would match ~90% of the Japanese population at the HLA-A, HLA-B and HLA-DRB1 loci with two-digit specification 12. We performed a similar estimation with four-digit specification using the HLA Laboratory database and found that 50 unique HLA-homozygous donors would cover ~73% of the Japanese population (Fig. 3a and Supplementary Table 8). Approximately 75 and 140 unique donors would be needed to cover ~80% and 90%, respectively. It would be necessary to type ~37,000, ~64,000 and ~160,000 individuals, respectively, to identify these 50, 75 and 140 donors (Fig. 3b). Allografts using HLA-homozygous iPSCs may provide a therapeutic alternative to autologous grafts, for cases in which transplant is likely to be needed soon after injury; furthermore, they allow for the advance selection of safe clones 13. The beneficial effects of matching at major HLA loci are well documented in renal transplantation 14,15 , although recipients of allografts derived from HLA-homozygous iPSCs would still need immunosuppressants after transplantation because of other HLA antigens, non-HLA antigens and immunity by natural killer cells. We report a simple, non-integrative method for reprogramming human cells. The increased efficiency and the use of nontransforming Myc should be useful to generate iPSCs from many donors, such as individuals with disease. The approach may also prove beneficial for generating human iPSCs for use in autologous and allologous stem cell therapy. Accession codes.