Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Exp Med. Nov 20, 2016; 6(4): 72-79
Published online Nov 20, 2016. doi: 10.5493/wjem.v6.i4.72
Morphogenesis of human embryonic stem cells into mature neurons under in vitro culture conditions
Geeta Shroff
Geeta Shroff, Nutech Mediworld, New Delhi 110016, India
Author contributions: Shroff G conceived and designed the study, and acquired, analyzed and interpreted the data. The author drafted the article and made critical revisions related to the intellectual content of the manuscript, and approved the final version of the article to be published.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board of Nutech Mediworld, New Delhi, India.
Informed consent statement: Not applicable.
Conflict-of-interest statement: The author declares no conflict of interest regarding the publication of this paper.
Data sharing statement: Our all publications are PubMed indexed and are available online (open access). We shall not be applying for membership of the Dryad Repository.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Correspondence to: Geeta Shroff, MBBS, Director, Nutech Mediworld, H-8, Green park extension, New Delhi 110016, India.
Telephone: +91-11-26180039 Fax: +91-11-26560089
Received: March 24, 2016
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.

Keywords: Human embryonic stem cells, Multipotency, Neural differentiation, Neural progenitor cells, In-vitro fertilization

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.