RNA-Seq of Single Lung Circulating Tumor Cells (CTCs) and primary Tumor Cells from an orthotopic lung xenograft model

An orthotopic lung xenograft model was used to identify differential expression of lung CTCs in comparison to primary lung tumor cells. Overall design: Briefly, 300,000 MGH134 (a patient-derived non-small cell lung cancer line [Pubmed ID 25758528]) cells stably expressing luciferase and GFP were introduced into the lung by percutaneous injection. Mice blood samples were collected 7-8 weeks after injection, and CTCs were isolated using the CTC iChip [Pubmed ID 23552373]. Primary tumors were digested into single cells by collagenase and hyaluronidase (Stemcell Technologies). A total of 10 GFP-positive single CTCs and 6 single primary tumor cells were individually micromanipulated using a 10 µm transfer tip on an Eppendorf TransferMan NK 2 micromanipulator. Single cell sequencing was performed under the same procedure as described in [Pubmed ID 26383955].