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Culture of HCT116 cell 3D spheroidal tumor

SA Saloni S. Andhari
RW Ravindra D. Wavhale
KD Kshama D. Dhobale
BT Bhausaheb V. Tawade
GC Govind P. Chate
YP Yuvraj N. Patil
JK Jayant J. Khandare
SB Shashwat S. Banerjee
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3D tumor spheroids were formed by a modified method of the hanging drop technique49. In brief, the lid of sterile 12 well plates were coated with poly(dimethoxysiloxane) (PDMS) and Sylgard 184 in a 10:1 ratio and cured at 80 °C for around 45 min. The lids were then placed under UV for 30 min to ensure sterility of the PDMS coated surface. HCT116 cell suspension was prepared in complete McCoy’s 5A medium. 20 μL drops of the cell suspension with a density of 2,500 cells/drop were placed at regular intervals on the PDMS coated lid. The wells were filled with sterile MilliQ water to ensure hydration of drops upon incubation. Thereafter, the cells were incubated at 37 °C in presence of 5% CO2 for three days. Finally, the coherent mass of 3D tumor spheroids formed was selected for further studies.

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