One-Step Fabrication of Low-Cost, Autoclavable, and Multifunctional Silk-Based Nanofibrous Permeable Hanging Cell Culture Inserts for Various Biological Applications

[Image: see text] Hanging cell culture inserts are most widely used in vitro cell culture devices, which provide a freestanding multichamber setup for various co-culture and triculture systems. Apart from being costly, the commercial inserts do not provide enough choices regarding polymer types and pore sizes. Most importantly, commercially available inserts are two-dimensional multiporous membrane-based devices. Herein, we report a one-step fabrication process of the multifunctional nanofiber-based permeable hanging cell culture insert using electrospinning. These fabricated nanofibrous membranes’ attached inserts have advantages such as low cost, ready availability, easy fabrication, tunable porosity, autoclavability, and biomaterial-based nanofibrous membranes. The inserts without nanofibrous membrane can also be reused by autoclaving them and electrospun nanofibrous membrane on it according to the application. We have also confirmed its suitability for extensive use in the field of in vitro cell culture by analyzing its adherence and toxicity results on breast cancer cell line (MCF-7). These hanging cell culture inserts are thus a potent product for various cell culture assays such as cell migration for wound healing, cancer metastasis, and other tissue engineering applications.