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A 96-Well Plate Clonogenic Cell Assay for Squamous Cell Carcinoma

Reidar Grenman, MD; David Burk, MS; Erkki Virolainen, MD; John G. Wagner, PhD; Allen S. Lichter, MD; Thomas E. Carey, PhD

Arch Otolaryngol Head Neck Surg. 1988;114(4):427-431.

Abstract
• Radiation sensitivity was determined for nine University of Michigan squamous cell carcinoma (UM-SCC) cell lines, MCF-7 and HeLa, using a 96-well plate clonogenic assay. Plating efficiencies (PE) of the UM-SCC cell lines were between 0.16 and 0.36. Higher PE values obtained were with MCF-7 (0.4) and HeLa (0.5). The UM-SCC cell lines were used at low passages (passage 13 to passage 20) to minimize artifacts attributable to long-term culture. Cells were irradiated in suspension using a cobalt 60 gamma source at a dose rate of 0.94 Gy/min (94 rad/min). Survival data were fitted well by either a linear quadratic function F = e^–(D+βD2) or by a monoexponential function F = Ae^–D. Mean inactivation dose, equivalent to the area under the survival curve (AUC), was used as a measure of radiation sensitivity. The UM-SCC-1, 9, 11A, 11B, MCF-7, and HeLa were the most radiation resistant lines we tested (AUC >2.1), while UM-SCC-14A was the most sensitive (AUC = 1.591). The assay was highly reproducible, and the differences in radiation sensitivity between cell lines were statistically significant.

(Arch Otolaryngol Head Neck Surg 1988;114:427-431)

Author Affiliations
From the Cancer Research Laboratory of the Department of Otolaryngology/Head and Neck Surgery (Mr Burk and Dr Carey), the Departments of Radiation Oncology (Dr Lichter) and Pharmacology (Dr Wagner), and the College of Pharmacy (Dr Wagner), University of Michigan, Ann Arbor; the Department of Otorhinolaryngology, Turku (Finland) University Hospital (Dr Virolainen); and Loimaa (Finland) Regional Hospital (Dr Grenman).

Footnotes
Accepted for publication Dec 14, 1987.

Reprint requests to Cancer Research Laboratory, 6020 Kresge Hearing Research Institute, Box 0506, 1301 E Ann St, University of Michigan, Ann Arbor, MI 48109-0506 (Dr Carey).

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