title: Survival and engraftment of dopaminergic neurons manufactured by a Good Manufacturing Practice-compatible process.
abstract: OBJECTIVE
  We have previously reported a Good Manufacturing Practice (GMP)-compatible process for generating authentic dopaminergic neurons in defined media from human pluripotent stem cells and determined the time point at which dopaminergic precursors/neurons (day 14 after neuronal stem cell [NSC] stage) can be frozen, shipped and thawed without compromising their viability and ability to mature in vitro. One important issue we wished to address is whether dopaminergic precursors/neurons manufactured by our GMP-compatible process can be cryopreserved and engrafted in animal Parkinson disease (PD) models.
  
  
  METHODS
  In this study, we evaluated the efficacy of freshly prepared and cryopreserved dopaminergic neurons in the 6-hydroxydopamine-lesioned rat PD model.
  
  
  RESULTS
  We showed functional recovery up to 6 months post-transplantation in rats transplanted with our cells, whether freshly prepared or cryopreserved. In contrast, no motor improvement was observed in two control groups receiving either medium or cells at a slightly earlier stage (day 10 after NSC stage). Histologic analysis at the end point of the study (6 months post-transplantation) showed robust long-term survival of donor-derived tyrosine hydroxylase (TH)(+) dopaminergic neurons in rats transplanted with day 14 dopaminergic neurons. Moreover, TH(+) fibers emanated from the graft core into the surrounding host striatum. Consistent with the behavioral analysis, no or few TH(+) neurons were detected in animals receiving day 10 cells, although human cells were present in the graft. Importantly, no tumors were detected in any grafted rats, but long-term tumorigenic studies will need to determine the safety of our products.
  
  
  CONCLUSIONS
  Dopaminergic neurons manufactured by a GMP-compatible process from human ESC survived and engrafted efficiently in the 6-OHDA PD rat model.
authors: Buck Institute for Age Research, Novato, California, USA.
authors: Jun
authors: J
authors: Peng
authors: Buck Institute for Age Research, Novato, California, USA.
authors: Qiuyue
authors: Q
authors: Liu
authors: NxCell, Novato, California, USA.
authors: Mahendra S
authors: MS
authors: Rao
authors: Buck Institute for Age Research, Novato, California, USA; XCell Science, Novato, California, USA. Electronic address: xzeng@buckinstitute.org.
authors: Xianmin
authors: X
authors: Zeng
mesh_terms: D000818:Animals; D001709:Biotechnology; D002460:Cell Line; D002470:Cell Survival; D003342:Corpus Striatum / Q000473:pathology; D015925:Cryopreservation / Q000379:methods*; D004195:Disease Models, Animal; D059290:Dopaminergic Neurons / Q000473:pathology / Q000502:physiology*; D005260:Female; D006801:Humans; D066192:Manufacturing Industry; D009412:Nerve Fibers / Q000473:pathology; D058953:Neural Stem Cells / Q000473:pathology / Q000502:physiology*; D055495:Neurogenesis; D016627:Oxidopamine / Q000008:administration & dosage / Q000009:adverse effects; D010300:Parkinson Disease / Q000209:etiology / Q000628:therapy*; D017410:Practice Guidelines as Topic; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function; D033581:Stem Cell Transplantation; D014183:Transplantation, Heterologous
chemical_list: D016627:Oxidopamine
keywords: GMP; cell therapy; dopaminergic neuron; human pluripotent stem cells