Conditionally immortalized stem cell lines from human spinal cord retain regional identity and generate functional V2a interneurons and motorneurons

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• 作者：Graham Cocks (1)
  Nataliya Romanyuk (2)
  Takashi Amemori (2)
  Pavla Jendelova (2) (3)
  Oksana Forostyak (2)
  Aaron R Jeffries (1)
  Leo Perfect (1)
  Sandrine Thuret (1)
  Govindan Dayanithi (2) (4)
  Eva Sykova (2) (3)
  Jack Price (1)
  
• 关键词：Neural stem cells ; Spinal cord ; V2a interneurons ; Motoneurons ; Voltage ; operated Ca2+ channels ; Spontaneous Ca2+ oscillations
• 刊名：Stem Cell Research & Therapy
• 出版年：2013
• 出版时间：September 2013
• 年：2013
• 卷：4
• 期：3
• 全文大小：
• 作者单位：Graham Cocks (1)
  Nataliya Romanyuk (2)
  Takashi Amemori (2)
  Pavla Jendelova (2) (3)
  Oksana Forostyak (2)
  Aaron R Jeffries (1)
  Leo Perfect (1)
  Sandrine Thuret (1)
  Govindan Dayanithi (2) (4)
  Eva Sykova (2) (3)
  Jack Price (1)
  
  1. The James Black Centre, Department of Neuroscience, King’s College London, 125 Coldharbour Lane, London, UK
  2. Institute of Experimental Medicine, ASCR, Prague, Czech Republic
  3. Department of Neuroscience, 2nd Faculty of Medicine, Charles University, Prague, Czech Republic
  4. Institut National de la Santé et de la Recherche Médicale, Unité de recherche U710, Montpellier and Ecole Pratique des Hautes Etudes, Université Montpellier 2, Paris, F-75007, France
  
• ISSN：1757-6512

文摘

Introduction The use of immortalized neural stem cells either as models of neural development in vitro or as cellular therapies in central nervous system (CNS) disorders has been controversial. This controversy has centered on the capacity of immortalized cells to retain characteristic features of the progenitor cells resident in the tissue of origin from which they were derived, and the potential for tumorogenicity as a result of immortalization. Here, we report the generation of conditionally immortalized neural stem cell lines from human fetal spinal cord tissue, which addresses these issues. Methods Clonal neural stem cell lines were derived from 10-week-old human fetal spinal cord and conditionally immortalized with an inducible form of cMyc. The derived lines were karyotyped, transcriptionally profiled by microarray, and assessed against a panel of spinal cord progenitor markers with immunocytochemistry. In addition, the lines were differentiated and assessed for the presence of neuronal fate markers and functional calcium channels. Finally, a clonal line expressing eGFP was grafted into lesioned rat spinal cord and assessed for survival, differentiation characteristics, and tumorogenicity. Results We demonstrate that these clonal lines (a) retain a clear transcriptional signature of ventral spinal cord progenitors and a normal karyotype after extensive propagation in vitro, (b) differentiate into relevant ventral neuronal subtypes with functional T-, L-, N-, and P/Q-type Ca2+ channels and spontaneous calcium oscillations, and (c) stably engraft into lesioned rat spinal cord without tumorogenicity. Conclusions We propose that these cells represent a useful tool both for the in vitro study of differentiation into ventral spinal cord neuronal subtypes, and for examining the potential of conditionally immortalized neural stem cells to facilitate functional recovery after spinal cord injury or disease.