Monodisperse semi-permeable microcapsules for continuous observation of cells

Yuya Morimoto, Wei-heong Tan, Yukiko Tsuda, Shoji Takeuchi
2009 Lab on a Chip  
Supplementary information: Formation of microcages with mammalian cells 1. Materials, solution preparation and cell culture In addition to materials cited in the main text, Dulbecco's modified eagle's medium (DMEM), DMEM powder and Trypsin-EDTA solution were purchased from SIGMA-Aldrich. Fetal bovine serum (FBS) was purchased from Japan Bioserum Co. Ltd.. LIVE/DEAD® Viability/Cytotoxicity kit for Live/Dead assay was purchased from Invitrogen Co.. Mammalian adherent cells used here were 3T3
more » ... here were 3T3 cells that are mouse fibroblast cells. 3T3 cells were cultured in DMEM supplemented with 10 v/v% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin at 37°C under a humidified atmosphere of 5% CO 2 . Cell viability was examined with LIVE/DEAD ® Viability/Cytotoxicity kit following manufacturer's protocol. DMEM (without NaHCO 3 ) solution was prepared as follows: 2.7 g DMEM powder was added to 200 ml of sterilized water and pH was subsequently adjusted to 7.1 with NaOH. Once a homogeneous solution was obtained, the solution was sterilized by filtration. Methods of producing microcages with mammalian adherent cells For producing microcages with mammalian adherent cells, we modified the gelation method using GDL and CaCO 3 . In the AFFD, corn oil with 2 wt.% lecithin was used as the outer fluid. For the inner fluid, DMEM (without NaHCO 3 ) with GDL (10 mM) and DMEM (without NaHCO 3 ) with sodium alginate (2 wt.%) containing insoluble CaCO 3 (75 mg/ml) and 3T3 cells were used, respectively. When we used DMEM instead of DMEM (without NaHCO 3 ) for producing sodium alginate solution, ph reduction with carbonate ions in DMEM induced the gelation of alginate as calcium ions were released from calcium nano particles. Therefore, we cannot produce monodisperse alginate sol droplets. The inner fluids to produce monodisperse alginate sol droplets with cells were
doi:10.1039/b900035f pmid:19606299 fatcat:vw6y7y6ucjgplbmvvcwouxhjku