Cell culture tests

Biocompatibility test

Date	1st June 2020
Short Description	Evaluating possible toxicity and other negative effects of the perfusion system and its components on endothelial cell culture.
Passing Criteria	Cell proliferation and metabolic activity are comparable to a control sample. The perfusion system features allow improvements above the control sample: A larger volume of medium supply, reducing required amount of manual work. Introduction of medium flow and other parameters for cell culture research.
Details	A basic perfusion set-up was assembled to test basic biocompatibility: a tray module silicone tubing luer-lock tubing connectors (PP and stainless steel) 20ml syringes (PP) medium flow manifold (POM) 6-well plate breathable well membrane The set-up was assembled as shown in the Figures below and a culture of human umbilical vein endothelial cells (HUVEC) was introduced into two wells, a testing sample (connected to flow) and a control sample (static). The set-up was installed into a standard cell culture incubator and maintained for 1 week at 37°C and 5% CO2 with intermittent replenishment of culturing medium in the control sample.
Results	The biocompatibility experiments have produced the following results: The breathable well membrane allows sufficient gas exchange for long term experiments The POM, tubing and connectors do not produce toxic effects Continuous use of single use syringes results in wear of the plastic, increasing pumping resistance and producing plastic debris, which enters the cell culture. Thus, single use syringes need to be exchanged every 24h and are less suitable for long-term use.

HUVEC cells after 1h.

HUVEC cells after 24h.

HUVEC cells after 72h.