Substrates and supplements for hESCs: a critical review

Crocco, M.; Fratnz, Nilo; Bös-Mikich, Adriana

doi:10.1007/s10815-012-9914-8

Cited by 14 publications

(11 citation statements)

References 80 publications

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“…One of the major challenges in the development of a derivation system for clinical-grade hESC lines has been the lack of a suitable combination of culture medium with supporting feeder cells or feeder-free matrix to support expansion of the inner cell mass (ICM) [ 28 ]. We initially used human placental stromal fibroblasts (hPSFs) to replace mouse embryonic fibroblasts (MEFs) as feeder cells with xeno-free knock-out serum (KO-SR ZF) hESC medium [ 29 ].…”

Section: Resultsmentioning

confidence: 99%

High quality clinical grade human embryonic stem cell lines derived from fresh discarded embryos

Bates

Soteriou

et al. 2017

Stem Cell Res Ther

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BackgroundHuman embryonic stem cells (hESCs) hold tremendous promise for cell replacement therapies for a range of degenerative diseases. In order to provide cost-effective treatments affordable by public health systems, HLA-matched allogeneic tissue banks of the highest quality clinical-grade hESCs will be required. However only a small number of existing hESC lines are suitable for clinical use; they are limited by moral and ethical concerns and none of them apply Good Manufacturing Practice (GMP) standards to the earliest and critical stages of gamete and embryo procurement. We thus aimed to derive new clinical grade hESC lines of highest quality from fresh surplus GMP grade human embryos.MethodsA comprehensive screen was performed for suitable combinations of culture media with supporting feeder cells or feeder-free matrix, at different stages, to support expansion of the inner cell mass and to establish new hESC lines.ResultsWe developed a novel two-step and sequential media system of clinical-grade hESC derivation and successfully generated seven new hESC lines of widely varying HLA type, carefully screened for genetic health, from human embryos donated under the highest ethical and moral standards under an integrated GMP system which extends from hESC banking all the way back to gamete and embryo procurement.ConclusionsThe present study, for the first time, reports the successful derivation of highest-quality clinical-grade hESC lines from fresh poor-quality surplus human embryos generated in a GMP-grade IVF laboratory. The availability of hESC lines of this status represents an important step towards more widespread application of regenerative medicine therapies.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0561-y) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

High quality clinical grade human embryonic stem cell lines derived from fresh discarded embryos

Bates

Soteriou

et al. 2017

Stem Cell Res Ther

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“…These include deriving cells from oocytes unsuitable for IVF following parthenogenetic activation (De Sousa et al 2009;Camarasa et al 2012), from growth-arrested IVF embryos (Zhang et al 2006), and from otherwise nonviable or poor-quality embryos (Gavrilov et al 2011;Lerou et al 2008;Ye et al 2017). Conventionally, the separated, expanded ICM-derived cells are then cultured in medium which promotes pluripotent stem cell growth, on a substrate formed of a lawn of non-mitotic "feeder" cells, such as mitotically inactivated mouse embryonic fibroblasts (MEFs) (Thomson et al 1998;Crocco et al 2013). Islands of cells emerge with the classical ESC morphology of small cells with a high nucleus to cytoplasm ratio and prominent nucleoli.…”

Section: Derivationmentioning

confidence: 99%

“…Initially the provision of the, then unknown, signaling factors was facilitated by growing colonies of hESCs on a lawn of non-mitotic MEFs. The dependence on the use of MEFs was problematic as they are of nonhuman origin, and the composition of signaling and extracellular proteins that they secrete and present can be highly variable between batches and labs (Lim and Bodnar 2002;Chin et al 2007;Crocco et al 2013). Despite many advances in substrates and media, there is not yet a robust, universally accepted feeder-free protocol for the derivation and culture of karyotypically normal hESC lines (International Stem Cell Initiative et al 2011).…”

Section: Substratesmentioning

confidence: 99%

“…Despite many advances in substrates and media, there is not yet a robust, universally accepted feeder-free protocol for the derivation and culture of karyotypically normal hESC lines (International Stem Cell Initiative et al 2011). In order to avoid the potential contamination of these cultures with animal-derived pathogens, a wide variety of xeno-free feeder cells have been developed such as human dermal, foreskin, placental, or embryonic fibroblasts (HDFs, HFFs, HPFs, or HEFs) (Crocco et al 2013;Mckay et al 2011). These cell lines have been demonstrated to reliably replace MEFs, removing our reliance on them when deriving hESC lines.…”

Section: Substratesmentioning

confidence: 99%

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Embryonic Stem Cells

Lewis¹,

Silajdžić²,

Brison³

et al. 2017

Cell Engineering and Regeneration

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Embryonic stem cells (ESCs) are pluripotent stem cells derived from a preimplantation embryo. ESCs are distinguished by two major properties: their pluripotency (the ability to differentiate into all derivatives of the three primary germ layers) and their ability to replicate indefinitely under defined conditions. Human ESCs (hESCs) can be used to study early human development and genetic disease and for in vitro toxicology testing. Because of their plasticity and potentially unlimited capacity for self-renewal, clinical-grade hESC therapies have been proposed for tissue replacement after injury or disease. In this chapter we summarize the process of hESC derivation, discuss characterization (the standard tests that are performed during the cell culture process to check that the cells exhibit the fundamental properties that make them ESCs), and provide examples of protocols that are used to induce hESCs to differentiate into specific cell types.

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“…Despite the encouraging results obtained using the synthetic gel formulations and the existence of clinical‐grade human embryonic stem cell lines (Ilic et al, ), there is the need to find easily manufactured, safe, and economical materials for use as three‐dimensional (3D) substrates for pluripotent stem cells (Crocco et al, ).…”

Section: Introductionmentioning

confidence: 99%

Interaction of fibroblasts and induced pluripotent stem cells with poly(vinyl alcohol)‐based hydrogel substrates

et al. 2019

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Induced pluripotent stem cells (iPSCs) provide a promising means of creating custom‐tailored cell lines for cellular therapies. Their application in regenerative medicine, however, depends on the possibility that the maintenance and differentiation of cells and organs occur under defined conditions. One major component of stem cell culture systems is the substrate, where the cells must attach and proliferate. The present study aimed to investigate the putative cytotoxic effects of poly(vinyl alcohol) (PVA)‐based matrices on the in vitro culture of mouse fetal fibroblasts. In addition, the PVA‐based hydrogels were used to determine the capacity of bovine induced pluripotent stem cells (biPSCs) to adhere and proliferate on synthetic substrates. Our results show that both cell types interacted with the substrate and presented proliferation during culture. The biPSCs formed new colonies when cell suspensions were placed onto the hydrogel surface for culture. These results may represent a new characterized xeno‐free clinical grade culture system to be widely applied in cell‐based therapies.

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Substrates and supplements for hESCs: a critical review

Abstract: This information shall be useful for the creation and choice of new substrates and supplements for future research in the field of hESC for therapeutic purposes.

Cited by 14 publications

References 80 publications

High quality clinical grade human embryonic stem cell lines derived from fresh discarded embryos

High quality clinical grade human embryonic stem cell lines derived from fresh discarded embryos

Embryonic Stem Cells

Interaction of fibroblasts and induced pluripotent stem cells with poly(vinyl alcohol)‐based hydrogel substrates

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