Parkinson’s disease (PD) is a common neurodegenerative condition characterized by progressive and selective neuronal cell loss. Banks of cryopreserved committed mDA neural progenitor cells provide a method to generate human DA neurons with reduced batch-to-batch variability, and establish a mechanism to share lineage-primed cells for collaborative research. Importantly, cryopreservation of mDA neural progenitor cells did not alter their potential to resume differentiation into mDA neurons. A faster cooling rate of 1–2☌/min was significantly better than 0.5☌/min for all conditions tested, while rapid thawing at 37☌ was not always superior to slow thawing at 4☌. The presence of ROCK inhibitors improved cell recovery at 24 h for all cryopreservation media tested. Significant differences in cell recovery were observed at 24 h post-thawing, but no differences were observed immediately upon thawing. Here we have compared six (6) different commercial cryopreservation media and different freezing conditions for mDA neural progenitor cells differentiated from human embryonic stem cell (hESC) lines. Creating a bank of cryopreserved mDA neural progenitor cells poised for neuronal differentiation could significantly improve reproducibility and facilitate collaborations. One solution to this problem is to cryopreserve committed mDA neural progenitor cells in a ready-to-use format. However, due to the complexity and length of the protocols and inherent differences between cell lines, considerable variability of the final population of neurons is often observed. Combined with the unlimited growth potential of pluripotent stem cells, mDA neural progenitor cell production can provide a scalable source of human dopaminergic (DA) neurons for diverse applications. The developmental identity of mDA neurons produced using floor plate protocols is similar to substantia nigra neurons, and this has improved the ability to model Parkinson’s disease (PD) in a dish.
Human pluripotent stem cells can be differentiated into midbrain dopaminergic (mDA) neurons by directing cells through a floor plate progenitor stage. 3UK Centre for Mammalian Synthetic Biology, The University of Edinburgh, Edinburgh, United Kingdom.1MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, United Kingdom.Drummond 1, Karamjit Singh Dolt 1, Maurice A.
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