Figure 5.

IL89_CL#3488 continuous cell line generation and characterization. (A) Schematic IL89_CL#3488 continuous cell line derivation strategy. The patient’s material was digested and cultured overnight. Floating lymphoma cells were isolated and cultured alone or with stromal elements, attached to the plate. (B) Growth curve of IL89 cells cultured alone or in the presence of stromal elements. (C) TCR gamma specific rearrangement clonality of IL89 PDTX T5 and the IL89_CL#3488 continuous cell line showed the identity of the tumor clone. (D) Heatmap of the most significantly mutated genes in IL89 detected by WES shows a similarity between IL89 T5 and the IL89_CL#3488 continuous cell line. The color-code provided is indicative of the VAF. (E) RNA expression levels of a breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) gene signature shows a strict association between the IL89 diagnostic sample, PDTX (T1-T3-T5), and the IL89_CL#3488 continuous cell line. (F) Unsupervised hierarchical clustering of total RNA sequencing profiles of IL89 (PDTX and cell line) vs, IL17 ALK-ALCL model. Samples of the same models cluster together. (G) Principal component analysis based on total RNA sequencing data of IL89 (Primary-PDTX-cell line) vs. IL17 models showed that the expression landscapes of the primary tumors were maintained in the respective PDTX models and that the two different entities have distinct transcriptomes. The IL89_CL#3488 cell line fits in the same cluster as its primary and PDTX-derived passages. (H) IL89_CL#3488 recapitulates the ruxolitinib sensitivity of the corresponding PDTC (72 h, 1 µM). Data are representative of at least three independent experiments, and values are expressed as the average ± standard deviation. p-values were calculated using the student t-test. ** p < 0.01, *** p < 0.001.