An ECM-based culture system for the generation and maintenance of xeno-free human iPS cells

Kim, Hyeong-Taek; Lee, Kang-In; Kim, Dong-Wook; Hwang, Deuk-Soo

doi:10.1016/j.biomaterials.2012.10.064

Cited by 29 publications

(15 citation statements)

References 32 publications

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“…However, in terms of cell reprogramming, they reported that the iPSC colony formation efficiencies of fibroblasts on iMattix511 ranged from 0.02% to 0.034% (28), which is lower than the 0.13% efficiency obtained on the Matrigel system in our study. Consistent with their study and other previous studies (3,5,8), we showed that the feeder systems based on MEFs and FLCs were much more efficient in iPSC generation than the feeder-free Matrigel system, providing the efficiencies ranging from 1.25% to 14.7%. Thus, while feeder-free systems are valuable for generating clinical-grade hiPSCs for cell transplantation and can be used to generate hiPSCs with growth and differentiation potential equivalent to feeder-grown hiPSCs, further improvement of cell programming efficiency on a feeder-free system would add more value and be useful for iPSC generation from certain types of cells that are difficult to reprogram.…”

Section: Discussionsupporting

confidence: 94%

Human iPS cell-derived fibroblast-like cells as feeder layers for iPS cell derivation and expansion

Tay

Chen

et al. 2015

Journal of Bioscience and Bioengineering

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Section: Discussionsupporting

confidence: 94%

Human iPS cell-derived fibroblast-like cells as feeder layers for iPS cell derivation and expansion

Tay

Chen

et al. 2015

Journal of Bioscience and Bioengineering

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“…Using vitronectin and xeno-free differentiation culture medium, the hiPSCs were able to differentiate into motor neurons under xeno-free conditions, and expressed markers of motor neurons. This xeno-free culture system greatly reduces the risk of immune rejection after cell transplantation and provides an invaluable tool for drug discovery and regenerative medicine by generation of patient-specific and clinical-grade iPSCs [20,28].…”

Section: Discussionmentioning

confidence: 99%

“…Extraction of genomic DNA and bisulfite mutagenesis sequencing analysis were conducted using a ReadyAmp Genomic Kit (Promega, Madison, WI) and EZ DNA Methylation Kit (Zymo Research, Orange, CA), respectively, according to the manufacturers' instructions. After bisulfite mutation, the DNA was eluted in 20-μl elution buffer and subjected to 35 cycles of PCR with primer pairs for the OCT4 promoter (forward 5′-ATT TGTTTT TTGGGTAGTTAAAGGT-3′ and reverse 5′-CCAACTATCT TCATCTTAA TAACATCC-3′) and NANOG promoter (forward 5′-AGAGATAGGAGGG TAAGTT TT TTTT-3′ and reverse 5′-ACTCCCACACAAACTAACTTTTATTC-3′) [28]. PCR products were verified by 2 % agarose gel electrophoresis.…”

Section: Bisulfite Genomic Sequencingmentioning

confidence: 99%

Derivation, Expansion, and Motor Neuron Differentiation of Human-Induced Pluripotent Stem Cells with Non-Integrating Episomal Vectors and a Defined Xenogeneic-free Culture System

Xiong

et al. 2015

Mol Neurobiol

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Induced pluripotent stem cells (iPSCs) generated from patient-derived somatic cells provides the opportunity for model development in order to study patient-specific disease states with the potential for drug discovery. However, use of lentivirus and exposure of iPSCs to animal-derived products limit their therapeutic utility and affect lineage differentiation and subsequent downstream functionality of iPSC derivatives. Within the context of this study, we describe a simple and practical protocol enabling the efficient reprogramming of terminally differentiated adult fibroblasts into integration-free human iPSCs (hiPSCs) using a combination of episomal plasmids with small molecules (SMs). Using this approach, there was a 10-fold increase in reprogramming efficiency over single plasmid vector-based methods. We obtained approximately 100 iPSCs colonies from 1 × 10(5) human adult dermal fibroblasts (HADFs) and achieved approximately 0.1% reprogramming efficiencies. Concurrently, we developed a highly conducive culture system using xeno-free media and human vitronectin. The resulting hiPSCs were free of DNA integration and had completely lost episomal vectors, maintained long-term self-renewal, featured a normal karyotype, expressed pluripotent stem cell markers, and possessed the capability of differentiating into components of all three germ layers in vivo. Finally, we demonstrate that the integration-free hiPSCs could be differentiated into motor neurons under xeno-free culture conditions. This induction method will promote the derivation of patient-specific integration-free and xeno-free iPSCs and improve the strategy for motor neuron derivation. Our approach provides a useful tool for human disease models, drug screen, and clinical applications.

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“…Methods have been described for the expansion of pluripotent stem cells using completely defined, xeno-free cell culture medium and attachment matrices (Chen et al, 2011; Kim et al, 2013; Rajala et al, 2010; Wang et al, 2012). hiPSCs are expanded a predetermined amount based on the intended use of the bank, taking into consideration the disease indication and expected future need.…”

Section: The Path To Hipsc Clinical Trials: Manufacturing Clinicalmentioning

confidence: 99%

Induced pluripotent stem cells as custom therapeutics for retinal repair: Progress and rationale

Wright

Phillips

Pinilla

et al. 2014

Experimental Eye Research

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Human pluripotent stem cells have made a remarkable impact on science, technology and medicine by providing a potentially unlimited source of human cells for basic research and clinical applications. In recent years, knowledge gained from the study of human embryonic stem cells and mammalian somatic cell reprogramming has led to the routine production of human induced pluripotent stem cells (hiPSCs) in laboratories worldwide. hiPSCs show promise for use in transplantation, high throughput drug screening, “disease-in-a-dish” modeling, disease gene discovery, and gene therapy testing. This review will focus on the first application, beginning with a discussion of methods for producing retinal lineage cells that are lost in inherited and acquired forms of retinal degenerative disease. The selection of appropriate hiPSC-derived donor cell type(s) for transplantation will be discussed, as will the caveats and prerequisite steps to formulating a clinical Good Manufacturing Practice (cGMP) product for clinical trials.

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An ECM-based culture system for the generation and maintenance of xeno-free human iPS cells

Cited by 29 publications

References 32 publications

Human iPS cell-derived fibroblast-like cells as feeder layers for iPS cell derivation and expansion

Human iPS cell-derived fibroblast-like cells as feeder layers for iPS cell derivation and expansion

Derivation, Expansion, and Motor Neuron Differentiation of Human-Induced Pluripotent Stem Cells with Non-Integrating Episomal Vectors and a Defined Xenogeneic-free Culture System

Induced pluripotent stem cells as custom therapeutics for retinal repair: Progress and rationale

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