Author = سقا، محسن

Isolation and primary culture of chick embryonic neural crest cells

Volume 11, Issue 4, Winter 2021, Pages 275-282

https://doi.org/10.52547/JCT.11.4.275

M Matin, MGH Golmohammadi, M Sagha

Abstract Aim: we aimed at presenting a simple and efficient method for isolating and characterizing the neural crest cells.
Material and Methods: The hen’s fertilized eggs were incubated for about 35h at 38°c and 55-60% humidity until the embryos reached to stages 10-12 according to Hamburger-Hamilton developmental stage table. Then the embryos were removed from the egg’s yolk and the neural tube was isolated and cultured for 24 h in a tissue culture dish to release neural crest cell. Then after, the neural tube was removed and allowed to NCC to expand for further 5 days. Finally, the cells were collected and subjected to PCR to study their gene expression profile.
Results: The neural tube released NCC and these cells proliferated in culture condition. They also expressed markers including Slug, Sox9 ,and Sox10 by the RT-PCR method.
Conclusion: The neural tube can release NCC in culture condition and these cells can proliferate in the presence of an appropriate medium.
 

Investigating the effect of Cirsium vulgare hydroethanolic extract on neural stem cells proliferation

Volume 5, Issue 4, Winter 2015, Pages 385-391

https://doi.org/10.52547/JCT.5.4.385

A A, SM Kh, M S, F S, D N, R B

Abstract Aim: In vitro investigation of hydroethanolic extraction of Cirsium vulgare on growth rates of neonate rat neural stem cells. Material and Methods: Neural stem cells were isolated from hippocampus of neonatal rat brain. To determine optimal concentration of Cirsium vulgare extraction, isolated neural stem cells were treated with 200, 400, 600, 800 and 1000 µg/ml for 48 h and then cells proliferation rate were evaluated by MTT assay. In addition Sox2 mRNA expression in neural stem cells was evaluated using quantitative real-time PCR. Results: The results of this study showed that the Cirsium vulgare extract caused significant increase in cell proliferative activity and Sox2 mRNA expression when compared with the control group. Conclusion: with respect to the effect of Cirsium vulgare extract on the proliferation rate of neural stem cells, the use of hydroethanolic extraction of Cirsium vulgare can be used for clinical purposes to treat some of the neurodegenerative disorders such as ischemic stroke and spinal cord injury.

Sclerotomal Differentiation of Somitic Cells Co-Cultured with Chicken Embryonic Notochord

Volume 5, Issue 1, Summer 2014, Pages 71-77

https://doi.org/10.52547/JCT.5.1.71

R R, M M, MGh G, M S

Abstract Aim: The somites are epithelial blocks of mesodermal cells differentiating to sclerotome and dermomyotome and notochord or axial mesoderm plays a major role in somitic cell differentiation to sclerotome. This study was aimed to evaluate the role of the notochord in sclerotomal differentiation of the somites in vitro. Material and Methods: In this experimental study, isolated notochords from chick embryo were encapsulated in alginate beads and co-cultured with somites for six days. In control group, the somites were cultured alone. Newly isolated and non-cultured somites were considered as somites on day 0. Eventually, morphology of cultured somitic cells was evaluated by inverted microscope and then RT-PCR method was used to analyze the sclerotomal and dermomyotomal gene expression profile. Results: In comparison with control group, the somitic cells co-cultured with notochord had highly differentiation potential and showed mesenchymal morphology with numerous slender processes. Gene expression profile of co-cultured somitic cells indicated upregulation of Pax1 and BMP4 and downregulation of MyoD, presenting sclerotomal cell characteristics. Conclusion: Embryonic notochord can induce in vitro sclerotomal differentiation in somites through upregulation of genes involving in this differentiation.