Navigating challenges: optimising methods for primary cell culture isolation

• Published in: Cancer cell international Vol. 24; no. 1; p. 28
• Main Authors: Piwocka, Oliwia, Musielak, Marika, Ampuła, Karolina, Piotrowski, Igor, Adamczyk, Beata, Fundowicz, Magdalena, Suchorska, Wiktoria Maria, Malicki, Julian
• Format: Journal Article
• Language: English
• Published: England BioMed Central 11.01.2024; BMC
• Subjects: Biopsy; Breast cancer; Cancer cells; Cancer therapies; Cancer-associated fibroblasts; Cell culture; Collagenase; Cytokeratin; Disaggregation; Enzymatic digestion; Epidermal growth factor; Fibroblasts; Isolation; Methods; Mortality; Patients; Penicillin; Primary culture; France; United States > US; Cancer-associated fibroblasts; Cancer cells; Enzymatic digestion; Primary culture; Isolation; Breast cancer; Primary cell line
• ISSN: 1475-2867; 1475-2867
• DOI: 10.1186/s12935-023-03190-4

Primary cell lines are invaluable for exploring cancer biology and investigating novel treatments. Despite their numerous advantages, primary cultures are laborious to obtain and maintain in culture. Hence, established cell lines are still more common. This study aimed to evaluate a range of techniques for isolating primary breast cancer cultures, employing distinct enzymatic compositions, incubation durations, and mechanical approaches, including filtration. Out of several protocols, we opted for a highly effective method (Method 5) that gave rise to a primary cell culture (BC160). This method combines mechanical disaggregation and enzymatic digestion with hyaluronidase and collagenase. Moreover, the paper addresses common issues in isolating primary cultures, shedding light on the struggle against fibroblasts overgrowing cancer cell populations. To make primary cell lines a preferred model, it is essential to elaborate and categorise isolation methods, develop approaches to separate heterogeneous cultures and investigate factors influencing the establishment of primary cell lines.