Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells

Sun, Ninghui; Panetta, Nicholas J.; Gupta, Deepak; Wilson, Kitchener D.; Lee, Andrew; Jia, Fangjun; Hu, Shijun; Cherry, Athena M.; Robbins, Robert C.; Longaker, Michael T.; Wu, Joseph C.

doi:10.1073/pnas.0908450106

Cited by 461 publications

(349 citation statements)

References 29 publications

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“…Lentivirus Production and Transduction-Lentivirus for reprogramming transgenes or Fluc-RFP-HSVttk (firefly luciferase, red fluorescent protein, and herpes simplex truncated thymidine kinase) was collected from the supernatant of transfected 293FT cells and concentrated as described previously (11,12).…”

Section: Adipose Cell Harvest and Culture Of Canine Ipscs (Cipscs)-mentioning

confidence: 99%

Preclinical Derivation and Imaging of Autologously Transplanted Canine Induced Pluripotent Stem Cells

Lee

Plews

et al. 2011

Journal of Biological Chemistry

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110

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Derivation of patient-specific induced pluripotent stem cells (iPSCs) opens a new avenue for future applications of regenerative medicine. However, before iPSCs can be used in a clinical setting, it is critical to validate their in vivo fate following autologous transplantation. Thus far, preclinical studies have been limited to small animals and have yet to be conducted in large animals that are physiologically more similar to humans. In this study, we report the first autologous transplantation of iPSCs in a large animal model through the generation of canine iPSCs (ciPSCs) from the canine adipose stromal cells and canine fibroblasts of adult mongrel dogs. We confirmed pluripotency of ciPSCs using the following techniques: (i) immunostaining and quantitative PCR for the presence of pluripotent and germ layer-specific markers in differentiated ciPSCs; (ii) microarray analysis that demonstrates similar gene expression profiles between ciPSCs and canine embryonic stem cells; (iii) teratoma formation assays; and (iv) karyotyping for genomic stability. Fate of ciPSCs autologously transplanted to the canine heart was tracked in vivo using clinical positron emission tomography, computed tomography, and magnetic resonance imaging. To demonstrate clinical potential of ciPSCs to treat models of injury, we generated endothelial cells (ciPSC-ECs) and used these cells to treat immunodeficient murine models of myocardial infarction and hindlimb ischemia.

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Section: Adipose Cell Harvest and Culture Of Canine Ipscs (Cipscs)-mentioning

confidence: 99%

Preclinical Derivation and Imaging of Autologously Transplanted Canine Induced Pluripotent Stem Cells

Lee

Plews

et al. 2011

Journal of Biological Chemistry

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110

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“…iPS cells are usually maintained on feeder cells that are derived from mouse embryonic fibroblasts (MEFs) (Takahashi and Yamanaka 2006;Takahashi et al 2007;Yu et al 2007). However, several feederfree conditions were developed recently especially for human embryonic stem (ES) cells and iPS cells to ensure biosafe human clinical applications (Xu et al 2001;Amit et al 2004;Rosler et al 2004;Lu et al 2006;Navarro-Alvarez et al 2008;Miyazaki et al 2008;Sun et al 2009;Kitajima and Niwa 2010;Rodin et al 2010;Hayashi et al 2010;Fukusumi et al 2013). Animal-derived components are relatively high risk for potential infections in clinical regenerative medicine.…”

Section: Introductionmentioning

confidence: 99%

Low-molecular-weight inhibitors of cell differentiation enable efficient growth of mouse iPS cells under feeder-free conditions

Donai

Inagaki

et al. 2014

Cytotechnology

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Embryonic stem cells and induced pluripotent stem (iPS) cells are usually maintained on feeder cells derived from mouse embryonic fibroblasts (MEFs). In recent years, the cell culture of iPS cells under serum-and feeder-free conditions is gaining attention in overcoming the biosafety issues for clinical applications. In this study, we report on the use of multiple small-molecular inhibitors (i.e., CHIR99021, PD0325901, and Thiazovivin) to efficiently cultivate mouse iPS cells without feeder cells in a chemically-defined and serum-free condition. In this condition, we showed that mouse iPS cells are expressing the Nanog, Oct3/4, and SSEA-1 pluripotent markers, indicating that the culture condition is optimized to maintain the pluripotent status of iPS cells. Without these small-molecular inhibitors, mouse iPS cells required the adaptation period to start the stable cell proliferation. The application of these inhibitors enabled us the shortcut culture method for the cellular adaptation. This study will be useful to efficiently establish mouse iPS cell lines without MEF-derived feeder cells.

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“…induced pluripotent stem cells (hiPSCs) have the potential to grow indefinitely in culture and to make any cell in the adult human body, they represent important resources for regenerative medicine (1) and for disease modeling efforts that seek to recreate a patient's disease pathogenesis in the laboratory (2,3). hESCs and hiPSCs are typically grown on a "feeder" cell layer of mitotically inactivated mouse embryonic fibroblasts (mEFs) or on "feeder-free" culture systems composed of a variety of ECM/ serum proteins coated onto tissue culture dishes (4)(5)(6)(7)(8)(9) or synthetic materials (10)(11)(12)(13)(14). Nearly all these systems have been reported to promote hESC/hiPSC self-renewal when the cells are passaged in multicellular clumps and seeded at a suitably high cell density.…”

mentioning

confidence: 99%

Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions

Saha

Mei²,

Reisterer³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

135

109

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The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.

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Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells

Cited by 461 publications

References 29 publications

Preclinical Derivation and Imaging of Autologously Transplanted Canine Induced Pluripotent Stem Cells

Preclinical Derivation and Imaging of Autologously Transplanted Canine Induced Pluripotent Stem Cells

Low-molecular-weight inhibitors of cell differentiation enable efficient growth of mouse iPS cells under feeder-free conditions

Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions

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