Karbalaie Khadijeh, Tanhaei Somayyeh, Rabiei Farzaneh, Kiani-Esfahani Abbas, Masoudi Najmeh Sadat, Nasr-Esfahani Mohammad Hossein, Baharvand Hossein
Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Cell J. 2015 Spring;17(1):37-48. doi: 10.22074/cellj.2015.510. Epub 2015 Apr 8.
The neural crest is a transient structure of early vertebrate embryos that generates neural crest cells (NCCs). These cells can migrate throughout the body and produce a diverse array of mature tissue types. Due to the ethical and technical problems surrounding the isolation of these early human embryo cells, researchers have focused on in vitro studies to produce NCCs and increase their knowledge of neural crest development.
In this experimental study, we cultured human embryonic stem cells (hESCs) on stromal stem cells from human exfoliated deciduous teeth (SHED) for a two-week period. We used different approaches to characterize these differentiated cells as neural precursor cells (NPCs) and NCCs.
In the first co-culture week, hESCs appeared as crater-like structures with marginal rosettes. NPCs derived from these structures expressed the early neural crest marker p75 in addition to numerous other genes associated with neural crest induction such as SNAIL, SLUG, PTX3 and SOX9. Flow cytometry analysis showed 70% of the cells were AP2/P75 positive. Moreover, the cells were able to self-renew, sustain multipotent differentiation potential, and readily form neurospheres in suspension culture.
SHED, as an adult stem cell with a neural crest origin, has stromal-derived inducing activity (SDIA) and can be used as an NCC inducer from hESCs. These cells provide an invaluable resource to study neural crest differentiation in both normal and disordered human neural crest development.
神经嵴是早期脊椎动物胚胎中的一种短暂结构，可产生神经嵴细胞（NCCs）。这些细胞能够迁移至全身并产生多种成熟的组织类型。由于围绕这些早期人类胚胎细胞分离存在的伦理和技术问题，研究人员专注于体外研究以产生NCCs并增进他们对神经嵴发育的了解。
在这项实验研究中，我们将人类胚胎干细胞（hESCs）与人脱落乳牙的基质干细胞（SHED）共培养两周。我们采用不同方法将这些分化细胞鉴定为神经前体细胞（NPCs）和NCCs。
在共培养的第一周，hESCs呈现为带有边缘玫瑰花结的火山口样结构。源自这些结构的NPCs除了表达与神经嵴诱导相关的众多其他基因如SNAIL、SLUG、PTX3和SOX9外，还表达早期神经嵴标志物p75。流式细胞术分析显示70%的细胞AP2/P75呈阳性。此外，这些细胞能够自我更新，维持多能分化潜能，并易于在悬浮培养中形成神经球。
SHED作为一种起源于神经嵴的成体干细胞，具有基质衍生诱导活性（SDIA），可作为从hESCs诱导产生NCCs的诱导剂。这些细胞为研究正常和紊乱的人类神经嵴发育中的神经嵴分化提供了宝贵资源。
