Uwe Rix, Kenneth E. Huffman, John D. Minna, Eric B. Haura, Tingting Jiang, Oliver A. Hampton, Jamie K. Teer, Boning Gao, Vasiliki Pelekanou, Harsha Doddapaneni, Fumi Kinose, Joy Jayaseelan, Robert C. Doebele, Jennifer R. Peters-Hall, Kemp H. Kernstine, Wei Zhang, Luc Girard, Yuval Kluger, Dara L. Aisner, Melissa Coquelin, David A. Wheeler, Chunxian Huang, Marileila Varella-Garcia, Kyle R. Covington, Dwight H Oliver, Anh T. Le, David L. Rimm, Jerry W. Shay, Adi F. Gazdar, and Jianhong Hu
// Boning Gao 1, 2, 3 , Chunxian Huang 1, 2 , Kemp Kernstine 4 , Vasiliki Pelekanou 5 , Yuval Kluger 5 , Tingting Jiang 6 , Jennifer R. Peters-Hall 7 , Melissa Coquelin 7 , Luc Girard 1, 2, 3 , Wei Zhang 1, 2 , Kenneth Huffman 1, 2 , Dwight Oliver 8 , Fumi Kinose 9 , Eric Haura 9 , Jamie K. Teer 10 , Uwe Rix 11 , Anh T. Le 12 , Dara L. Aisner 13 , Marileila Varella-Garcia 12, 13 , Robert C. Doebele 12 , Kyle R. Covington 14 , Oliver A. Hampton 14 , Harsha V. Doddapaneni 14 , Joy C. Jayaseelan 14 , Jianhong Hu 14 , David A. Wheeler 14 , Jerry W. Shay 7 , David L. Rimm 5 , Adi Gazdar 1, 2, 8 , John D. Minna 1, 2, 3, 15 1 Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA 2 The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA 3 Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA 4 Division of Thoracic Surgery, University of Texas Southwestern Medical Center, Dallas, Texas, USA 5 Department of Pathology, Yale University, New Haven, CT, USA 6 Interdepartmental Program in Computational Biology and Bioinformatics, Yale University, New Haven, CT, USA 7 Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA 8 Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas, USA 9 Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA 10 Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA 11 Department of Drug Discovery, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA 12 Department of Medicine, Division of Medical Oncology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA 13 Department of Pathology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA 14 Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA 15 Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA Correspondence to: Adi Gazdar, email: Adi.Gazdar@UTSouthwestern.edu John D. Minna, email: John.Minna@UTSouthwestern.edu Keywords: conditionally reprogrammed cells, respiratory epithelial cells, non-small cell lung cancer, rock inhibitor, cell culture Received: September 23, 2016 Accepted: December 20, 2016 Published: December 29, 2016 ABSTRACT The “conditionally reprogrammed cells” (CRC) method, using a Rho kinase inhibitor and irradiated mouse fibroblast cells has been described for the efficient growth of cells from malignant and non-malignant samples from primary tumor and non-malignant sites. Using the CRC method, four institutions independently cultured tumor tissues from 48 non-small cell lung cancers (NSCLC, mostly from primary resected tumors) and 22 non-malignant lungs. We found that epithelial cells could be cultured from tumor and non-malignant lung. However, epithelial cells cultured from tumors had features of non-malignant respiratory epithelial cells which include: 1) among 22 mutations found in the original tumors only two mutations were found in the CRC cultures with reduced frequency (31% to 13% and 92% to 15% from original tumor and CRC culture respectively); 2) copy number variation was analyzed in 9 tumor and their CRC cultures and only diploid patterns were found in CRC cultures; 3) mRNA expression profiles were similar to those of normal respiratory epithelial cells; and 4) co-culture of tumor and non-malignant lung epithelial cells resulted in mostly non-malignant cells. We conclude that CRC method is a highly selective and useful method for the growth of non-malignant respiratory epithelial cells from tumor specimens and only occasionally do such CRC cultures contain a small subpopulation of cancer cells marked by oncogenic mutations. While our findings are restricted to resected primary NSCLC, they indicated the necessity to fully characterize all CRC cultures and the need to develop culture technology that facilitates the growth of primary lung cancers.