Title: Twelve-day medium pumping into tubular cell-laden scaffold using a lab-made PDMS connector

Authors: V-T Duong, T-T Dang, JP Kim, K Kim, H Ko, CH Hwang, K-i Koo

Address: Department of Biomedical Engineering, University of Ulsan, Ulsan, Republic of Korea. room 409, building 18, University of Ulsan, 93 Daehak-ro, Nam-gu, Ulsan (44610), Republic of Korea

E-mail: kikoo at ulsan.ac.kr

Abstract: In the current study, a method is proposed to supply culture medium into a two-layered cell-laden tubular scaffold in order to enhance cell proliferation, confluence, and viability. The two-layered cell-laden tubular scaffold was made of calcium-alginate mixed with fibroblast cells (NIH/3T3) using a lab-made double- coaxial laminar-flow generator. Afterwards, the tubular scaffold was connected to a syringe pump system using a polydimethylsiloxane (PDMS) micro-connector for long-term cell culture. Three medium pumping conditions were applied and compared: a heart-beat-mimicking pumping (20 µL/s, 1 s period, and 50 % pulse width), a continuous pumping (20 µL/s) and a non-pumping. Non-leaky connections between the tubular scaffolds and the micro-connector outlet were sustained for 13.5 ± 0.83 d in heartbeat-mimicking pumping and 11.8 ± 0.33 d in continuous pumping condition, due to the elasticity of the tubular scaffolds. Importantly, the two pumping conditions resulted in more cell proliferation, confluence, and viability than the non-pumping condition. Furthermore, analysis of newly-produced type-I collagen matrix indicated that the cells under the two pumping conditions formed a tissue-like structure. The proposed technique could further be applied to vascular co-culturing for vascular engineered tissue.

Key Words: Two-layered cell-laden, calcium-alginate, double-coaxial generator, fibroblast, long-term culturing, media pumping.

Publication date: July 23rd 2019

Article download: Pages 1-13 (PDF file)
DOI: 10.22203/eCM.v038a01