An optimized method for neuronal differentiation of embryonic stem cells in vitro.
BACKGROUND
Neural differentiation from embryonic stem cells (ESCs) is an excellent model for elucidating the key mechanisms involved in neurogenesis, and also provides an unlimited source of progenitors for cell-based nerve regeneration. However, the existing protocols such as small molecule substances, 3D matrix, co-culture technique and transgenic method, are complicated and difficult to operate, thus are limited by laboratory conditions. Looking for an easy-to-operate protocol with easily gained material and high induction efficiency has always been a hot issue in neuroscience research.


NEW METHODS
This paper established an optimized method for embryonic neurogenesis using a strategy of "combinatorial screening". In our study, the whole process of embryonic neurogenesis was divided into two phases, and the differentiation efficiency of seven experimental protocols in phase I and three protocols in phase II were systematically evaluated in A2lox and 129 ESCs.


RESULTS
In phase I differentiation, "2-day embryoid bodies formation + 6-day retinoic acid induction" (Phase I-protocol 3) could effectively induce the differentiation of ESCs into neural precursor cells (NPCs). Furthermore, in phase II, N2B27 medium II (Phase II-protocol 3) could better support the subsequent differentiation from NPCs into neurons.


COMPARISON WITH EXISTING METHOD(S)
Such a combinational method (phase I-protocol 3 and phase II-protocol 3) can realize embryonic neurogenesis with high efficiency, easy implementation and low-cost, and is suitable for promotion in most laboratories.


CONCLUSIONS
Through "combinatorial screening" strategy, we established an optimized method for embryonic neurogenesis in vitro, which is expected to be a powerful tool for neuroscience research.
['Li|Yuan|Y|', 'Mao|Xiang|X|', 'Zhou|Xianyi|X|', 'Su|Yuting|Y|', 'Zhou|Xiangyu|X|', 'Shi|Kaituo|K|', 'Zhao|Shasha|S|']
D000818:Animals D002454:Cell Differentiation D046508:Culture Techniques D058732:Embryoid Bodies D053595:Embryonic Stem Cells D051379:Mice D018345:Mice, Knockout D058953:Neural Stem Cells D055495:Neurogenesis D009488:Neurosciences
Embryonic stem cells Neural differentiation Embryoid body Retinoic acid N2B27 medium