Microfabrication of Bubbular Cavities in PDMS for Cell Sorting and Microcell Culture Applications

Characterization of cell seeding and specific capture of B cells in microbubble well arrays

“…The BSA solution does not enter the MB wells. MBs were primed with PBS buffer using the vacuum-assisted coating (VAC) technique previously described (Giang et al 2008). Briefly, ~20 μl of PBS buffer was place onto the chip surface and then subjected to negative pressure in a bench top vacuum chamber for 45 min.…”

“…Relatively large reagent volumes, long processing times, and the necessity to use many plates to assay for minority cell types or secreted soluble factors (e.g. cytokines, antibodies) are additional limitations that can be overcome using microfabricated systems (Love et al 2006; Giang et al 2008; Liberskit et al 2011). The attributes of a low cell culture volume, customizable surface chemistry, and the ability to fabricate high density micro-well arrays, are particularly advantageous for immune system research in which both single cell studies and interactions between B and T cells can be specifically probed (Waldmann 1979; Lanzavecchia 1985; Love et al 2006; Tangye et al 2012).…”

“…Recently, we introduced microbubble (MB) well array technology and demonstrated its use to sustain single and small cell cultures for extended periods of time (>10 days) (Giang et al 2008; Chandrasekaran et al 2011). Microbubbles are spherical cavities formed in polydimethylsiloxane (PDMS) using the gas expansion molding (GEM) process (Giang et al 2007, 2012).…”

“…In previous work we have shown that MB arrays can be used to dissect the heterogeneity of a cell sample at the single cell level (Giang et al 2012). MB wells have also been used to grow arrays of homogeneously sized microtumors (Giang et al 2008), to study the epidermal-mesenchymal transition process that is important in cancer metastasis (Chandrasekaran et al 2011), and to determine the clonogenic potential of samples enriched with cancer stem cells (Chandrasekaran and DeLouise 2011). Furthermore, we have demonstrated a MB well array based perfusion system for culturing multi-cellular tumor spheroids and showed that spheroid cells were more resistant to doxorubicin treatment than adherent cells (Agastin et al 2011).…”

Gather Round: In vitro tumor spheroids as improved models of in vivo tumors

“…Our group has developed a gas expansion molding (GEM) technique for generating microbubbles which are 80-150 μm diameter spherical compartments formed on polydimethylsiloxane (PDMS), a silicone based elastomeric polymer. We showed that arrays of spherical cavities formed on PDMS [7] using this technique can be used for culturing cancer cells as spheroids [8]. The unique geometry of microbubbles allows the cells to rapidly condition their microenvironment by secreting soluble factors to influence their function [9].…”