A study of in-vitro cell growth for bioreactor constructs

Abstract

Tissue engineering is, despite many advances, still a long way from its aim of being able to reproduce in-vitro fully functionalised, vascularised musculoskeletal tissues. Pharmacy and research groups are currently in urgent need of reliable, comparable in-vitro testing grounds to analyse and test new drugs in order to gather more knowledge about physiology and pathology of these tissues and to fight diseases of the musculoskeletal system. This research aims to study different cell culture methodologies using a bioreactor to mature and differentiate muscle and tendon tissues, engineered tissues from different cell sources. This will allow further study on the effects of electrical, mechanical stimuli in-vitro and should lead to an analysis of the effect of single and combined stimulation methods applied to the subject tissue, this will further allow the finding of optimised parameters to mature and functionalise engineered muscle and tendon tissues. Using fibroblast and myoblast cells as the starter cells in the cell culture dish, it was observed that cell-cell adherence occurred which helped to accelerate the growth of the cells in the culture dish. The resulting two dimensional tissue structure needed a period of about 30 days at 37 degree Celsius in the incubator to grow the sufficient number of cells to enable further study in three dimensional tissue structure. When the required number of cells had been reached, a 3D scaffold made of fibrin was added which acted as the extra cellular matrix needed to allow the cells to form a more complex functionalised tissue. This was done by mixing solutions of fibrinogen and thrombin with a small amount of aprotinin solution. The aprotinin solution helps in delaying cell digesting of the fibrin gel and leaves enough fibrin gel to act as the extra cellular matrix for the three dimensional tissue formation.

It was also observed that cell contamination occurred if the cells were grown to more than 60-70% of the culture plate surface area, so cells were needed to be harvested before reaching this point as they would begin to start emitting chemicals which inhibit further growth. It was also important to prevent the contamination of the equipment as this would inhibit the cell growth. This report therefore describes the procedure in the equipment sterilization, the effect of different solutions in the cell growth and contamination in the cell culture procedure, much research effort was placed on this particular aspect.

Date of Award	2022
Original language	English
Awarding Institution	University of Dundee
Supervisor	Robert Keatch (Supervisor) & Jan Bernd Vorstius (Supervisor)