Primary Patient-Derived Cancer Cells and Their Potential for Personalized Cancer Patient Care

Abstract

SummaryPersonalized cancer therapy is based on a patient’s tumor lineage, histopathology, expression analyses, and/or tumor DNA or RNA analysis. Here, we aim to develop an in vitro functional assay of a patient’s living cancer cells that could complement these approaches. We present methods for developing cell cultures from tumor biopsies and identify the types of samples and culture conditions associated with higher efficiency of model establishment. Toward the application of patient-derived cell cultures for personalized care, we established an immunofluorescence-based functional assay that quantifies cancer cell responses to targeted therapy in mixed cell cultures. Assaying patient-derived lung cancer cultures with this method showed promise in modeling patient response for diagnostic use. This platform should allow for the development of co-clinical trial studies to prospectively test the value of drug profiling on tumor-biopsy-derived cultures to direct patient care.

Expansion of airway basal epithelial cells from primary human non‐small cell lung cancer tumors

“…Studies that report normal epithelial cell expansion have tended to initiate culture from early‐stage primary NSCLC tumors, whereas studies in which cancer cells have been expanded have initiated culture using biopsies or effusions from patients with late‐stage, therapy‐resistant disease7, 8, 9 or from PDX models,24 which may be more aggressive and/or more adaptable to cell culture. This would be consistent with traditional NSCLC cell culture methods and PDX models, where patient models are more readily established from late‐stage, metastatic or therapy‐resistant disease than from early‐stage primary disease.…”

“…We have previously demonstrated that adult human lung fibroblasts do not support long‐term normal airway epithelial cell expansion in the same way as 3T3‐J2 cells17 so the use of adult and/or human feeder cells might favor tumor cell expansion. Relevant biological differences might also exist between mitotic inactivation using irradiation4, 9 rather than mitomycin C, as used here. Alternatively, different tissue acquisition methods,9 feeder cell‐conditioned medium,10, 27 low oxygen,10 harsh tissue digestion protocols26 and/or initially serum‐starving cells6 might help to select tumor cells by inducing differentiation or apoptosis in normal epithelial cells.…”

“…In non‐small cell lung cancer (NSCLC), a number of reports demonstrate successful primary tumor cell culture using fibroblast co‐culture and ROCK inhibition 7, 8, 9, 10. However, others have found that normal epithelial cells are preferentially expanded in these conditions 11, 12.…”

Evaluation of holographic imaging cytometer holomonitor M4® motility applications

“…Over the decades, the development of high throughput and quantitative approaches to these assays has resulted in their widespread adoption as reliable assays for cell motility (4,5). One such application is the assessment of motility, proliferation, and drug response within short term ex vivo cultures of human cancers-an emerging approach for developing personalized therapeutic strategies (6,7). However, for some applications, these approaches are limited due to their requirement for culturing cells in conditions that deviate from those of routine culture and their reliance on measuring population averages.…”

“…However, for some applications, these approaches are limited due to their requirement for culturing cells in conditions that deviate from those of routine culture and their reliance on measuring population averages. One such application is the assessment of motility, proliferation, and drug response within short term ex vivo cultures of human cancers-an emerging approach for developing personalized therapeutic strategies (6,7). For this approach, assessing the behaviors of each cell in a heterogeneous population shortly after explant, without sacrificing the integrity of the culture, is ideal.…”