Obtaining Primary Ovarian Cancer Cells from Solid Specimens: A Method to Culture Epithelial Ovarian Cancer Cells from Ovarian Tumor Specimens
Published: April 30, 2023
Abstract
Source: Pribyl, L. J. et al. Method for Obtaining Primary Ovarian Cancer Cells From Solid Specimens. J. Vis. Exp. (2014)
In this video, we describe a method for isolating and characterizing primary ovarian cancer cells from solid ovarian tumor tissue specimens. The epithelial cancer cells obtained using this protocol are suitable for understanding the processes that may lead to ovarian cancer initiation and development.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Tissue Processing Working under sterile conditions, transfer the samples onto a Petri dish (60 mm x 60 mm) containing 10 ml of fresh, ice-cold PBS and using a sterile razor blade, further cut into the smallest pieces possible (2 mm or less) (<…
Representative Results
Figure 1. Processing of clinical specimens. A) Solid specimen transferred onto a Petri dish containing 10 ml of fresh, ice-cold 1x PBS. B) Clinical specimens further cut into pieces about 2 mm in size Figure 2. S…
開示
The authors have nothing to disclose.
Materials
| 1x PBS, sterile | Invitrogen | 14190-144 | |
| Screw lid, polypropylene specimen container, sterile |
Thermo Scientific | ||
| DMEM medium, sterile | Invitrogen | 11995-065 | |
| Fetal bovine serum, sterile | Thermo Scientific Hyclone | SH30396.03 | |
| 100x Penicillin-streptomycin, liquid | Invitrogen | 15140-122 | |
| Dispase II, sterile | Roche | 04942 078 001 | |
| Small fine-tip forceps and scalpel blade, sterile |
Fisher Scientific | forceps: 08-953E, blades: 08-927-5D |
|
| Small dissecting scissors, sterile | Fisher Scientific | 08-945 | |
| 15 ml Conical capped tubes, sterile | BD Falcon | 352097 | |
| 50 ml Conical capped tubes, sterile | BD Falcon | 352027 | |
| 10 ml Serological pipette, sterile | Corning | 13-678-11E | |
| 6 ml Syringe plunger, sterile | Tyco Healthcare | 8881516911 | |
| Nylon filter (70 μm ) cell strainer, sterile |
BD Falcon | 352350 | |
| 5 cm Petri dish, sterile | Corning | 430166 | |
| 10 cm Petri dish, sterile | Corning | 430167 |
