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The effect of omega-3 on muscle and non-muscle cells on a 2D fibronectin and in 3D fibrin constructs


Journal of Applied Biotechnology & Bioengineering
Sozzi Simon,1 Souza Danilo,1 SidhuKarandeep,1 Garcia Victor,1 Tawil Bill1,2

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Abstract

Human Foreskin Fibroblasts (HFF-1) and L6 muscle cells were treated in several conditions to examine the proliferation and adhesion of cells using different extracellular matrix proteins with different concentrations. These proteins consist of Collagen & Fibronectin with concentrations of 5ug/ml and 10ug/ml respectfully. The experiment was conducted and focused on how HFF-1 and L6 cells proliferate in two different conditions over a 7-day period and the results were recorded on Days 1, 3 & 7 (n=9). Different concentrations of Fibronectin and Collagen altered the proliferation of HFF-1 cells over a 7-day period. Once it was identified that the best proliferation of HFF-1 and L6 was the Fibronectin 10ug/mL coating, we used the same protein matrix to study the effects of different concentrations of Generic Omega-3 in cell adhesion and proliferation. Throughout the study, it is observed that proliferation is not proportional to the Omega-3 concentration, meaning in this study 1 mg/ml had the lowest proliferation. In conclusion from the first study, higher concentrations of Generic Omega-3 decreases the proliferation of HFF-1 and L6 cells. This is a known characteristic as Omega-3 inhibits cell proliferation by prolonging the G1 phase but did not arrest the G0-to-G1 or G1-to-S transitions. The prolonged G1 phase in fad3b ESCs was probably induced by downregulation of Cdk4 expression via p21 upregulation. In comparison to the Generic product of Omega-3 we decided to incorporate the Synthetic substance of Omega-3 into the study. The overall goal of this experiment was to observe the proliferation and adhesion of HFF-1 and L6 cells in artificially created media containing pure Synthetic Omega-3 fatty acids. This experiment was repeated three times (n=9) to gain valuable conclusive data which could be used to design future experiments. An additional experiment was specifically designed to see how HFF-1 and L6 cells migrate through a thick 3-D layer of fibrin. Fibrin is a substance formed through catalytic conversion of coagulation constituents: fibrinogen and thrombin. This experiment was conducted, and the results were taken for Day 1 and Day 7. Our results remained consistent in comparison to prior studies. With the use of higher concentrations of Synthetic Omega-3 there was a decrease in proliferation for both HFF-1 and L6 cells.

Keywords

HFF-1 cells, L6 muscle cells, fibronectin, collagen, omega-3, 3D fibrin

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