Directed Differentiation of V3 Interneurons from Mouse Embryonic Stem Cells.
نویسندگان
چکیده
Excitatory commissural V3 interneurons (INs) of the ventral spinal cord have been shown to balance locomotor rhythm regularity and robustness in vivo. Unfortunately, due to the scarcity of these cells in the spinal cord, in vitro studies of dissociated V3 INs have yet to be reported. In this study, we developed an induction protocol for V3 INs from mouse embryonic stem cells. The effect of the concentration of a strong sonic hedgehog (Shh) agonist (smoothened agonist [SAG]) and retinoic acid (RA) on expression of progenitor p3 and postmitotic V3 IN transcription factor markers (ie, Nkx2.2 and Sim1) was examined. Cells were differentiated toward a more ventral fate by increasing the duration of SAG exposure from 4 days in a previously established motoneuron induction protocol to 6 days. At the end of the induction period, transcription factor expression was assessed using quantitative real-time polymerase chain reaction, immunocytochemistry, in situ hybridization, and flow cytometry. Lower concentrations of RA and a longer duration of SAG exposure led to increased levels of p3 and V3 marker expression. This novel induction protocol reveals the importance of Shh signaling duration in the dorsal-ventral patterning of the neural tube, and it provides a method to obtain V3 INs for future studies to allow better understanding their role in rewiring and regeneration after spinal cord injury.
منابع مشابه
Differentiation of Mouse Embryonic Stem Cell into Insulin-Secreting Cell
Purpose: Differentiation of mouse embryonic stem cells into Insulin secreting endocrine cells. Materials and Methods: In this study, Royan B1 mouse embryonic stem cell (derived from C57BL/6 mouse) were used. In directed differentiation method, embryonicstem cells after embryoid bodies formation were differentiated into insulin secreting cells. Nestin positive cells were obtained after culture ...
متن کاملDifferentiation of Mouse Embryonic Stem Cells into Hematopoietic Cells
Purpose: Differentiation of Mouse embryonicstem cells into Hematopoietic cells. Materials and Methods: In this study, we used EB formation system for Hematopoietic differentiation of mouse embryonic stem cell (Royan B1) in suspension culture. EBs cultured in medium with Hematopoietic inducer cytokines (SCF, TPO, GMCSF, IL3, Flt3 and EPO) .presence of hematopoietic differentiated cell assessed ...
متن کاملThe Effect of Astrocyte-Conditioned Medium (ACM) and Retinoic Acid on Neural Differentiation of Mouse Embryonic Stem Cells
Purpose: The aim of this research was to study the properties of factors secreted from astrocyte cells in suspension medium in direct differentiation of mouse embryonic stem cells into neural cells. Materials and Methods: Royan B1 mouse embryonic stem (ES) cells were used in this experiment. For differentiation of Es cells into the neural cells, the astrocyte-condition medium (ACM) of mouse fe...
متن کاملFunctional Concentrations of BMP4 on Differentiation of Mouse Embryonic Stem Cells to Primordial Germ Cells
متن کامل
Pancreatic Differentiation of Sox 17 Knock-in Mouse Embryonic Stem Cells in Vitro
The way to overcome current limitations in the generation of glucose-responsive insulin-producing cells is selective enrichment of the number of definitive endoderm (DE) progenitor cells. Sox17 is the marker of mesendoderm and definitive endoderm. The aim of the present research was to study the potential of Sox17 knock-in CGR8 mouse embryonic stem (ES) cells to differentiate into insulin produ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Stem cells and development
دوره 24 22 شماره
صفحات -
تاریخ انتشار 2015