Dodecafluoroheptanol: Oxygen reservoir for the culture of mouse melanoma B16 cells

•Biphasic culture of animal cells using aqueous culture medium and a fluorous alcohol.•Mouse melanoma B16 cells thrived at the interphase between the two layers.•Formation of cell sheet at the interphase.•The dissolved oxygen content of the culture medium in the presence of various amounts of fluorous alcohol was monitored.•The fluorous solvent layer supplied oxygen that was consumed in the medium. Mouse melanoma B16 cells thrived during culture at the interphase between the aqueous culture medium and the fluorous solvent layers. However, cell spreading was not observed. Cells were rounded and attached to each other forming a cell sheet. Cells consumed the dissolved oxygen in the aqueous medium during culture and the depleted amount of oxygen was replenished in the presence of dodecafluoroheptanol. This research demonstrated that the fluorous solvent layer could serve as an oxygen reservoir for biphasic animal cell culture.