Efficient assembly of rat hepatocyte spheroids for tissue engineering applications

Engineering liver tissue spheroids with inverted colloidal crystal scaffolds

“…The general strategy is to prevent cell-substrate interactions while maximizing cell-cell interactions. Typical methods include the hanging drop technique [8], continuous agitation of suspension culture in a rotary cell culture vessel [9] or a spinner flask [10], preparation of cell repulsive substrates [11], and entrapment within biologically inert 3D hydrogel matrices [12,13]. In these culture conditions, single cells spontaneously self-assemble and form a spheroid aggregate.…”

Adhesion contact dynamics of primary hepatocytes on poly(ethylene terephthalate) surface

“…A group has recently demonstrated that the association rate constant of fibroblasts with fibronectin coated glass is positively correlated with the fibronectin density on the surface [25]. It has been recently shown that oxygen supply is critical to the formation and differentiated morphology of multicellular aggregates or spheroids of primary hepatocytes within 24 h after inoculation [26]. In our current study, we focused on the study of non-aggregated hepatocytes on a planar surface within the first 2 h of inoculation.…”

Spontaneous formation of stably-attached and 3D-organized hepatocyte aggregates on oxygen-permeable polydimethylsiloxane membranes having 3D microstructures

“…Several methods have thus been developed in order to induce hepatocyte spheroids formation on various kinds of substrates such as non-adherent plastic substrates (Landry et al 1985), proteoglycan-coated dishes (Koide et al 1989), positivelycharged surfaces (Koide et al 1990;Hansen et al 1998;Tzanakakis et al 2001;Peshwa et al 1994;Sakai and Suzuki 1991), PDMS surfaces (Nakazawa et al 2009), or macroporous scaffolds (Ijima et al 1998;Glicklis et al 2004;Dvir-Ginzberg et al 2004). Hepatocyte spheroids were also formed under rotational conditions in spinner vessels (Wu et al 1996;Sakai et al 1992) or in the microcavities of either a polystyrene (Fukuda and Nakazawa 2005) or a PDMS chip (Nakazawa et al 2006). However, these methods present several drawbacks: as their formation is promoted on surfaces which provide a weak cell-adherent environment, the spheroids formed are not kept attached to the substrates but are floating, thus leading to a poor applicability in microfluidic devices or in microplate-based drug/chemical screening where easy separation of cells and culture medium is required.…”