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Updated: Aug 5, 2026

A Simple Hanging Drop Cell Culture Protocol for Generation of 3D Spheroids
Published on: May 6, 2011
Spheroids formation in large drops suspended in superhydrophobic paper cones
Omkar Mohapatra1, Maheshwar Gopu1, Rahail Ashraf2
1Department of Physics, Indian Institute of Science Education and Research Tirupati, Mangalam P.O., 517507 Tirupati, AP, India.
Abstract:
The utilization of 3D cell culture for spheroid formation holds significant implications in cancer research, contributing to a fundamental understanding of the disease and aiding drug development. Conventional methods such as the hanging drop technique and other alternatives encounter limitations due to smaller drop volumes, leading to nutrient starvation and restricted culture duration. In this study, we present a straightforward approach to creating superhydrophobic paper cones capable of accommodating large volumes of culture media drops. These paper cones have sterility, autoclavability, and bacterial repellent properties. Leveraging these attributes, we successfully generate large spheroids of ovarian cancer cells and, as a proof of concept, conduct drug screening to assess the impact of carboplatin. Thus, our method enables the preparation of flexible superhydrophobic surfaces for laboratory applications in an expeditious manner, exemplified here through spheroid formation and drug screening demonstrations.
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