Efficient and simple approach to in vitro culture of primary epithelial cancer cells.
Primary cancer cells constitute a favourable testing platform for in vitro research in oncology field as they reflect tumour state more accurately than the most commonly employed stable cell lines. Unfortunately, due to limited availability of material and difficulties with protocols validation, primary models are rarely implemented into laboratory practice.We have compared protocols for primary cultures, differing in media components and plate coatings. In terms of culture establishment, application of Geltrex® coating demonstrated equal efficiency to feeder layer (83% compared with 72% successfully established breast and 80% compared with 80% prostate tumour specimens), yet it was substantially less complicated and easier to validate. Both Geltrex® coating and tissue-specific primary cell medium were permanently required to successfully maintain primary epithelial prostate cancer cells (PEPCs) in culture. In case of primary epithelial breast cancer cells (PEBCs), collagen I coating enabled to obtain comparable number of passages to Geltrex® coating (P=0.438). Commercial primary cell media demonstrated lower efficiency than tissue-specific ones (PEPCs-5 compared with 8 and PEBCs-6 compared with 9 passages). Interestingly, both analysed tumour types were unsusceptible to induction of culture lifespan extension when transduced with SV40LT, BMI-1 or hEST2 genes, commonly applied as potential immortalizing agents.In conclusion, the approach based on extracellular matrix reconstitution and tissue-specific primary cell media is easy to validate and provides in vitro expansion sufficient for analytical purposes (approximately 8 passages). Therefore, it may facilitate implementation of hardly available experimental models for a variety of analyses.
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland karolina.janik@celther.com.|Department of Tumor Biology, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland.
Karolina
K
Janik
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.
Marta
M
Popeda
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland.
Joanna
J
Peciak
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland.
Kamila
K
Rosiak
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.
Maciej
M
Smolarz
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.
Cezary
C
Treda
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland.
Piotr
P
Rieske
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland.
Ewelina
E
Stoczynska-Fidelus
Research and Development Unit, Celther Polska Ltd., Milionowa 23, 93-193 Lodz, Poland.
Magdalena
M
Ksiazkiewicz
D000818:Animals; D001940:Breast / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology*; D005109:Extracellular Matrix / Q000378:metabolism / Q000473:pathology; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D041681:NIH 3T3 Cells; D011467:Prostate / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000166:cytology*
D024042:Collagen Type I; D011518:Proto-Oncogene Proteins; D019098:Telomerase
Rho kinase (ROCK) inhibitor; breast cancer; extracellular matrix; primary cancer cell cultures; prostate cancer