title: Morphogenesis of human embryonic stem cells into mature neurons underin vitroculture conditions.
abstract: OBJECTIVE
  To describe the morphogenesis of different neuronal cells from the human embryonic stem cell (hESC) line, SCT-N, underin vitroculture conditions.
  
  
  METHODS
  The directed neuronal cell line was produced from a single, spare, pre-implantation stage fertilized ovum that was obtained during a naturalin vitrofertilization process. The hESCs were cultured and maintained as per our proprietary in-house technology in a Good Manufacturing Practice, Good Laboratory Practice and Good Tissue Practice compliant laboratory. The cell line was derived and incubated in aerobic conditions. The cells were examined daily under a phase contrast microscope for their growth and differentiation.
  
  
  RESULTS
  Different neural progenitor cells (NPCs) and differentiating neurons were observed under the culture conditions. Multipotent NPCs differentiated into all three types of nervous system cells,i.e., neurons, oligodendrocytes and astrocytes. Small projections resembling neurites or dendrites, and protrusion coming out of the cells, were observed. Differentiating cells were observed at day 18 to 20. The differentiating neurons, neuronal bodies, axons, and neuronal tissue were observed on day 21 and day 30 of the culture. On day 25 and day 30, prominent neurons, axons and neuronal tissue were observed under phase contrast microscopy. 4', 6-diamidino-2-phenylindole staining also indicated the pattern of differentiating neurons, axonal structure and neuronal tissue.
  
  
  CONCLUSIONS
  This study describes the generation of different neuronal cells from an hESC line derived from biopsy of blastomeres at the two-cell cleavage stage from a discarded embryo.
authors.affiliation: Geeta Shroff, Nutech Mediworld, New Delhi 110016, India.
authors.forename: Geeta
authors.initials: G
authors.lastname: Shroff
keywords: Human embryonic stem cells; In-vitro fertilization; Multipotency; Neural differentiation; Neural progenitor cells