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
Eur Cell Mater. 2019 Jul 23;38:1-13. doi: 10.22203/eCM.v038a01.
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.
在当前的研究中，提出了一种向双层细胞负载管状支架中供应培养基的方法，以增强细胞增殖、融合和活力。双层细胞负载管状支架由海藻酸钠与成纤维细胞（NIH/3T3）混合制成，使用实验室制造的双同轴层流发生器。之后，通过聚二甲基硅氧烷（PDMS）微连接器将管状支架与注射器泵系统连接，以进行长期细胞培养。应用并比较了三种培养基泵送条件：模拟心跳的泵送（20 μL/s、1 s 周期和 50%脉冲宽度）、连续泵送（20 μL/s）和非泵送。由于管状支架的弹性，在模拟心跳的泵送和连续泵送条件下，管状支架和微连接器出口之间的无泄漏连接可持续 13.5 ± 0.83 d 和 11.8 ± 0.33 d。重要的是，与非泵送条件相比，两种泵送条件导致更多的细胞增殖、融合和活力。此外，对新产生的 I 型胶原基质的分析表明，在两种泵送条件下的细胞形成了类似组织的结构。所提出的技术可以进一步应用于血管共培养以用于血管工程组织。
