Abstract
The successful isolation and propagation of patient-derived keratinocytes from cervical lesions constitute a more appropriate model of cervical disease than traditional cervical cancer-derived cell lines such as SiHa and CaSki. Our aim was to streamline the growth of patient-obtained, cervical keratinocytes into a reproducible process. We performed an observational case series study with 60 women referred to colposcopy for a diagnostic biopsy. Main outcome measures were how many samples could be passaged at least once (n = 11), and where enough cells could be established, to precisely define their proliferation profile over time (n = 3). Altering cell culture conditions over those reported by other groups markedly improved outcomes. We were also successful in making freeze backs which could be resuscitated to successfully propagate multi-layered, organoids from cervical keratinocytes (n = 3). For best results, biopsy-intrinsic factors such as size and tissue digestion appear to be major variables. This seems to be the first systematic report with a well characterized and defined sample size, detailed protocol, and carefully assessed cell yield and performance. This research is particularly impactful for constituting a sample repository-on-demand for appropriate disease modelling and drug screening under the umbrella of personalized health.
Highlights
The most practical and ethical approach seems to be obtaining biopsy specimens from patient lesions, while the physician is already taking a sample for standard-of-care
There is no literature on generating three-dimensional organotypic raft cultures from patient-derived, naturally-infected cervical keratinocytes
It would seem feasible to test new antiviral agents against human papillomavirus (HPV) such as the ones developed by us[9] in organoid models. In this observational case series study, we developed a patient-based cell culture model using freshly excised, cervical biopsies as a more accurate alternative to traditional cervical cancer-derived cell lines such as SiHa and CaSki
Summary
The most practical and ethical approach seems to be obtaining biopsy specimens from patient lesions, while the physician is already taking a sample for standard-of-care. It would seem feasible to test new antiviral agents against HPV such as the ones developed by us[9] in organoid models In this observational case series study, we developed a patient-based cell culture model using freshly excised, cervical biopsies as a more accurate alternative to traditional cervical cancer-derived cell lines such as SiHa and CaSki. Success was defined on how many samples could be propagated at least once, and grown long enough to precisely define their proliferation profile over time and whether freeze backs could be made and resuscitated for additional experiments. Initial results based on published methods[1,2,3,4,5] were clearly improved by altering cell culture conditions, and biopsy-intrinsic factors such as size and tissue digestion appear to be major variables This seems to be the first systematic report with a clearly defined sample size, detailed protocol, carefully assessed cell yield and performance, and the ability to grow cervical organoids. To develop such an approach seemed meaningful for constituting a sample repository-on-demand for appropriate disease modelling and drug studies
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call