Surface modifications to polydimethylsiloxane substrate for stabilizing prolonged bone marrow stromal cell culture

Polydimethylsiloxane (PDMS) has been extensively used as a supporting material for studies of cell mechanobiology, cell micropatterning and microscale-cell analysis in microfluidic chips due to its numerous advantages, such as low cytotoxicity, ease of modification, inexpensive costs and biocompatibility. However, the innate hydrophobicity of PDMS often poses a problem for stable cell adhesion, seriously limiting its applicability for prolonged cell culture. UV exposure and protein coating are suboptimal solutions, while chemical surface functionalization is often associated with laborious procedures and producing environmental toxics. Plasma treatment can render a hydrophilic substrate by altering the surface chemistry, but such effect is often short-lived due to its tendency to hydrophobic recovery. Variation of physical properties of the substratum are known to influence cell behaviour. Nevertheless, the combination of varying PDMS substratum properties via base:curing agent ratio and plasma treatment to stabilize the long-term culture of bone marrow derived stromal cells (BMSCs) still remain poorly understood. In this study, we developed a protocol to maintain the hydrophilicity of the plasma-treated PDMS over a range of substratum properties. This study demonstrated that varying the substratum properties of PDMS can enhance the stability of BMSC culture for at least three weeks, while plasma treatment with or without additional collagen coating further enhanced such effect. The changes in the physical properties of PDMS have rendered difference in BMSCs adhesion, proliferation and in-vitro plasticity, thereby offering a simple and effective strategy for PDMS surface modification to enable long term cell analysis in PDMS-based culture platform.