Revista de bioanálisis y biomedicina

Niche Induces Dedifferentiation

Abstract

Puja Sarkar, Aditi Kapoor, Jay Narayan Yadav and Sudhir Kumar

Background: Niche plays an important role in deciding the fate of the cell. Three dimensional (3-D) cell culture systems can provide conditions for the cells to revert back to de-differentiated state. Here, egg white is used as a scaffold as well as nutritive medium for the 3-D culture, as it provides nutrients for the cells to grow without extra media. In order to compare the growth of spheroids, another model was also taken into account that is gelatin coated plate. The difference between these two models was that the egg white acted as scaffold and provided additional nutrients for the growth of spheroids, while gelatin provided conditions for low attachment only. Method: Low attachment plates were prepared using egg white and gelatin for the spheroids to grow on it. RNA from cells grown on both 2D and 3D culture condition were extracted using trizol method. The concentration of RNA was determined by repeated OD measurements of aliquots at a wavelength of 260 nm and c-DNA was constructed using Fermentas kit. Finally, gene expressions were checked by PCR using thermo cycler. Conclusion: The cells had grown into spheroids on egg white based plate rather than gelatin coated plate. Gene expression was found to be upregulated in cells grown on egg white. Thus, it was proved that the niche provided by the egg white has induced de-differentiation, which leads to the acquisition of a stem cell like properties.