A novel xeno-free and feeder-cell-free system for human pluripotent stem cell culture

Wang, Qihui; Mou, Xiaoning; Cao, Hui; Meng, Qingzhang; Ma, Yan; Han, Pengcheng; Jiang, Junjie; Zhang, Hao

doi:10.1007/s13238-012-2002-0

Cited by 21 publications

(13 citation statements)

References 27 publications

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“…Many researchers have developed methods for maintaining, differentiating, and even freezing hPSCs in xeno-free conditions. 5,[35][36][37][38][39][40] Generating somatic cell populations from hPSCs using xeno-free culture conditions is essential for not only regenerative therapies to eliminate introduction of potential pathogens from animal-derived components into patients, but also for in vitro model systems that are designed to emulate a human system. Matrigel and gelatin are not ideal substrates for scale-up and application of hPSC-based culture systems given that there is lot-to-lot variability in the composition of these products and these substrates are not suitable for human tissue engineering applications as a result of their xenogeneic origin.…”

Section: Discussionmentioning

confidence: 99%

Efficient Generation of Functional Epithelial and Epidermal Cells from Human Pluripotent Stem Cells Under Defined Conditions

Selekman

Grundl

Kolz

et al. 2013

Tissue Engineering Part C: Methods

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Human pluripotent stem cells (hPSCs) have an unparalleled potential to generate limitless quantities of any somatic cell type. However, current methods for producing populations of various somatic cell types from hPSCs are generally not standardized and typically incorporate undefined cell culture components often resulting in variable differentiation efficiencies and poor reproducibility. To address this, we have developed a defined approach for generating epithelial progenitor and epidermal cells from hPSCs. In doing so, we have identified an optimal starting cell density to maximize yield and maintain high purity of K18+/p63+ simple epithelial progenitors. In addition, we have shown that the use of synthetic, defined substrates in lieu of Matrigel and gelatin can successfully facilitate efficient epithelial differentiation, maintaining a high (>75%) purity of K14+/p63+ keratinocyte progenitor cells and at a two to threefold higher yield than a previously reported undefined differentiation method. These K14+/p63+ cells also exhibited a higher expansion potential compared to cells generated using an undefined differentiation protocol and were able to terminally differentiate and recapitulate an epidermal tissue architecture in vitro. In summary, we have demonstrated the production of populations of functional epithelial and epidermal cells from multiple hPSC lines using a new, completely defined differentiation strategy.

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Section: Discussionmentioning

confidence: 99%

Efficient Generation of Functional Epithelial and Epidermal Cells from Human Pluripotent Stem Cells Under Defined Conditions

Selekman

Grundl

Kolz

et al. 2013

Tissue Engineering Part C: Methods

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“…This xeno-free culture model was also successfully used for terminal differentiation of the hESCs in to keratinocytes for potential therapeutic application [ 67 ]. Human placenta–derived ECM has also been successfully used for hESC culture [ 68 ]. Cultures showed genetic stability after 40 passages and differentiation potential of the stem cells was retained.…”

Section: Reviewmentioning

confidence: 99%

Human embryonic stem cell cultivation: historical perspective and evolution of xeno-free culture systems

Desai

Rambhia

Gishto

2015

Reprod Biol Endocrinol

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Human embryonic stem cells (hESC) have emerged as attractive candidates for cell-based therapies that are capable of restoring lost cell and tissue function. These unique cells are able to self-renew indefinitely and have the capacity to differentiate in to all three germ layers (ectoderm, endoderm and mesoderm). Harnessing the power of these pluripotent stem cells could potentially offer new therapeutic treatment options for a variety of medical conditions. Since the initial derivation of hESC lines in 1998, tremendous headway has been made in better understanding stem cell biology and culture requirements for maintenance of pluripotency. The approval of the first clinical trials of hESC cells for treatment of spinal cord injury and macular degeneration in 2010 marked the beginning of a new era in regenerative medicine. Yet it was clearly recognized that the clinical utility of hESC transplantation was still limited by several challenges. One of the most immediate issues has been the exposure of stem cells to animal pathogens, during hESC derivation and during in vitro propagation. Initial culture protocols used co-culture with inactivated mouse fibroblast feeder (MEF) or human feeder layers with fetal bovine serum or alternatively serum replacement proteins to support stem cell proliferation. Most hESC lines currently in use have been exposed to animal products, thus carrying the risk of xeno-transmitted infections and immune reaction. This mini review provides a historic perspective on human embryonic stem cell culture and the evolution of new culture models. We highlight the challenges and advances being made towards the development of xeno-free culture systems suitable for therapeutic 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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A novel xeno-free and feeder-cell-free system for human pluripotent stem cell culture

Cited by 21 publications

References 27 publications

Efficient Generation of Functional Epithelial and Epidermal Cells from Human Pluripotent Stem Cells Under Defined Conditions

Efficient Generation of Functional Epithelial and Epidermal Cells from Human Pluripotent Stem Cells Under Defined Conditions

Human embryonic stem cell cultivation: historical perspective and evolution of xeno-free culture systems

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

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