Abstract
BackgroundA 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.MethodsWe 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.ResultsHere, 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.ConclusionsOverall, 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.
Highlights
A major challenge for the clinical use of human pluripotent stem cells is the development of safe, robust and controlled differentiation protocols
The stem cell revolution started with the isolation of human embryonic stem cells [1] followed by the arrival of induced pluripotent stem cells [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
Development of an efficient good manufacturing practice (GMP)-compatible protocol for long-term neuroepithelial stem cell progenitors (lt-NES) derivation In order to develop a GMP-compatible protocol for ltNES derivation, we used H9 Human embryonic stem cell (hESC) routinely cultured on a widely recognised and defined culture system based on recombinant vitronectin and Essential 8 (E8) [13]
Summary
A major challenge for the clinical use of human pluripotent stem cells is the development of safe, robust and controlled differentiation protocols. 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. Policies around raw materials for cell therapeutics are currently flexible, clinical trials regulations require for each reagent to be extensively risk-assessed and qualified [8] It is in this context that so called GMP-grade materials suitable for clinical use will facilitate clinical trial submission and will likely become standard in the field [9]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
