Published online Nov 20, 2016. doi: 10.5493/wjem.v6.i4.72
Peer-review started: March 24, 2016
First decision: May 16, 2016
Revised: May 25, 2016
Accepted: July 14, 2016
Article in press: July 16, 2016
Published online: November 20, 2016
To describe the morphogenesis of different neuronal cells from the human embryonic stem cell (hESC) line, SCT-N, under in vitro culture conditions.
The directed neuronal cell line was produced from a single, spare, pre-implantation stage fertilized ovum that was obtained during a natural in vitro fertilization 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.
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.
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.
Core tip: Human embryonic stem cells (hESCs) have the capability to regenerate and differentiate into a wide variety of cells. In the present study, we described the morphogenesis of different neuronal cells from an hESC line under in vitro culture conditions. The blastomeres were at the two-cell cleavage stage and were taken from a discarded embryo during an in vitro fertilization process. We showed that neuronal axons and tissues were generated by the joining of multiple cells that communicate and transfer signals to each other, thereby forming neuronal cells and tissue.