GMP-grade neural progenitor derivation and differentiation from clinical-grade human embryonic stem cells
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RESEARCH
Open Access
GMP-grade neural progenitor derivation and differentiation from clinical-grade human embryonic stem cells Loriana Vitillo1* , Catherine Durance1, Zoe Hewitt2, Harry Moore2, Austin Smith1,3 and Ludovic Vallier1
Abstract Background: A major challenge for the clinical use of human pluripotent stem cells is the development of safe, robust and controlled differentiation protocols. Adaptation of research protocols using reagents designated as research-only to those which are suitable for clinical use, often referred to as good manufacturing practice (GMP) reagents, is a crucial and laborious step in the translational pipeline. However, published protocols to assist this process remain very limited. Methods: We adapted research-grade protocols for the derivation and differentiation of long-term neuroepithelial stem cell progenitors (lt-NES) to GMP-grade reagents and factors suitable for clinical applications. We screened the robustness of the protocol with six clinical-grade hESC lines deposited in the UK Stem Cell Bank. Results: Here, we present a new GMP-compliant protocol to derive lt-NES, which are multipotent, bankable and karyotypically stable. This protocol resulted in robust and reproducible differentiation of several clinical-grade embryonic stem cells from which we derived lt-NES. Furthermore, GMP-derived lt-NES demonstrated a high neurogenic potential while retaining the ability to be redirected to several neuronal sub-types. Conclusions: Overall, we report the feasibility of derivation and differentiation of clinical-grade embryonic stem cell lines into lt-NES under GMP-compliant conditions. Our protocols could be used as a flexible tool to speed up translation-to-clinic of pluripotent stem cells for a variety of neurological therapies or regenerative medicine studies. Keywords: Pluripotent stem cells, hESCs, Neural progenitors, GMP, Cell therapy
Background The stem cell revolution started with the isolation of human embryonic stem cells (hESCs) [1] followed by the arrival of induced pluripotent stem cells (iPSCs) [2], and their differentiation to an ever-increasing number of cell types has led to the prospect of shifting medicine to a new paradigm based on cellular repair. Despite this enticing prospect, the number of clinical trials based on * Correspondence: [email protected] 1 Wellcome-MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre; Department of Surgery, University of Cambridge, Cambridge CB2 0AW, UK Full list of author information is available at the end of the article
human pluripotent stem cells (hPSCs) remains limited when compared to other cell types [3]. There is a consensus that hPSCs have a complex and distinct set of scientific, technical and regulatory bottlenecks that hamper their translation to clinical applications [4–7]. One hurdle is that the often-large lists of raw materials used in differentiation protocols are designated of researchgrade and were never intended for cell therapy applications. The developers of advanced therapeutic medicinal
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