Novel approach for long term cell culturing usingthe PDMS connector

Abstract

We proposed a method to supply culture medium into a two-layered cell-laden tubular scaffold in order to enhance cell proliferation. The two-layered cell-laden tubular scaffold was made of calcium-alginate mixed with fibroblast cells using a lab-made double-coaxial laminar flow generator. Afterward, the tubular scaffold was connected to a syringe pump system using a polydimethylsiloxane (PDMS) micro-connector for long-term cell culturing. Three medium pumping conditions were applied and compared: a heart-beat mimicking pumping (20 µL/sec, 1 second period, and 50% pulse width), a continuous pumping (20 µL/sec) and a non-pumping. secure connections between the tubular scaffolds and the micro-connector outlet have sustained for 13.5 ± 0.83 days in heart-mimic pumping and 11.8 ± 0.33 days in continuous pumping condition due to elasticity of the tubular scaffolds. Importantly, the two pumping conditions showed more cell proliferation, confluence, and viability than the non-pumping condition. Also, analysis of newly-produced type-I collagen matrix indicated that the cells in the two pumping conditions formed a tissue-like structure. The proposed technique could further be applied to vascular co-culturing for vascular engineered tissue.