Bovine adipose tissue derived stem cells bioengineering: isolation, characterization and differentiation studies

Abstract

There is a significant potential for stem cells to be exploited for regenerative medicine. Stem cells act as a repair system for the body by differentiating into specialised cells and replenishing cells in regenerative organs such as skin or intestinal tissues. Conventionally in the current cell therapy, the renewed, differentiated cells were mostly sourced form bone marrow mesenchymal stem cell (MSCs) yielding low number of cells and involved a painful procedure under general anaesthesia. In tissue engineering, bioprocessing stem cells are crucial thus, stem cells from adipose tissue were revealed as a new promising source of MSCs. Abundant source of stromal vascular fraction (SVF) needs a good bioprocessing tools for an optimize culture. Isolation of bovine adipose tissue derived stem cells (ASCs) are found to better perform in 0.075% collagenase Type 1 and agitated for 2 hours from the experiment carried out. The static cultures were best inoculated at 1.0 x 104 cells/ml in 6 well plate. The suspension cultures were cultured best in 200rpm, 5ml DMEM: F12 / MesenPRO RS™ at 1.0 x 104 cells/ml in 50ml tube. Suspension (3D) culture of sphere aggregates yield cells density of 1.07 x 106 ASCs upon culture compared to static (2D) culture of 5 x 105 ASCs on average. However the viability of static culture are found to be more superior to suspension culture at high volume, agitation and inoculation sizes suggesting that the parameter are an unfavourable environment to culture in 50ml tubes. Successful differentiation of osteoblast cells were evaluated via Fourier Transform Infra-Red (FTIR) and conventional Van Kossa staining. Simulation of suspension (3D) culture via Computational Fluid Dynamic (CFD) using Volume of Fluid (VOF) model assist in developing an optimised ASCs culture protocol vis-à-vis conventional static (2D) culturing in flask.