Abstract
Adherent A431, BHK-21, and C2C12 cells were cultured on a flexible interface formed between two immiscible liquid phases: (i) hydrophobic perfluorodecalin (PFD) and (ii) aqueous culture medium (DMEM). BHK-21 cells formed multicellular aggregates characterized by irregular shapes. A431, as well as C2C12 cells, grew as tight multicellular sheets of 3-D cells. Enhanced mass transfer and facilitated access of the cells to the O2 dissolved in PFD/DMEM by approx. 250 % and thereby increased the density of BHK-21 cells. Thus the liquid/liquid system is a simple, ready-to-use, and fully scalable (independent of vessel shapes); consequently it is a method for 3-D cultures of adherent animal cells in which the growth of anchorage-dependent cells is not limited by confluence effect.
Highlights
The commonly-used systems of adherent animal cells culture require solid surfaces of culture dishes
Adherent A431, BHK-21, and C2C12 cells were cultured on a flexible interface formed between two immiscible liquid phases: (i) hydrophobic perfluorodecalin (PFD) and (ii) aqueous culture medium (DMEM)
An example of the flexible liquid/liquid culture system containing the air-saturated liquid PFD and Dulbecco’s modified Eagle’s medium (DMEM) has been presented on Fig. 1
Summary
The commonly-used systems of adherent animal cells culture require solid surfaces of culture dishes. The application of such culture methods is limited by the cells grown under such conditions usually forming surface-attached monolayers (Ulloa-Montoya 2005; Thomson et al 2007). The cells might form multilayer colonies or aggregates. In such case, the O2 supply to every cell might be a limiting factor (Fassnacht and Portner 1999; Harrison et al 2007). The application of O2 rich fluids, i.e. hydrophobic liquid O2 carriers, might be an alternative to conventional aeration systems (Leung et al 1997; Radisic et al 2006). The lack of toxicity and negative side effects of liquid PFCs on living cells has been confirmed by in vitro experiments and in clinical investigations (Mattiasson and Adlercreutz 1987; King et al 1989; Krafft 2001; Lowe 2002; Pilarek and Szewczyk 2008; Castro and Briceno 2010; Pilarek et al 2012; Hillig et al 2013)
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