Modeling Ovarian Cancer Metastasis in Peritoneal Cavity Lining: A Method To Establish an In Vitro 3D Model of the Peritoneal Lining
Published: April 30, 2023
Abstract
Source: Peters, P. N. et al. Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity. J. Vis. Exp. (2015)
In this video, we demonstrate the development of an in vitro three-dimensional model of the lining of the peritoneal cavity. This organotypic culture can help to investigate ovarian cancer cell adhesion, invasion, and proliferation.
Protocol
1. Plating the Organotypic Culture Release NOF (normal omental fibroblasts ) from a 75 cm2 culture flask by rinsing with 10 ml of PBS followed by 3 ml of 0.25% trypsin/25 mM EDTA for no more than 5 min. Neutralize trypsin with at least 3 times the volume of full growth media. Transfer trypsinized cells into a 50 ml conical tube and spin down at 0.5 g x 3 min. Remove supernatant and bring cells back up in 5 ml of full growth media. Use a cell counter to count the cell…
Açıklamalar
The authors have nothing to disclose.
Materials
| PBS | Fisher Scientific | SH3001304 | |
| 15 cm culture dishes | BD Biosciences | 353025 | |
| DMEM with L-Glutamine | Corning | 10-013-CV | |
| MEM Vitamins | Corning | 25-020-Cl | |
| MEM Nonessential amino acids | Corning | 25-025-CI | |
| Penicillin-Streptomycin | Corning | 30-002-CI | |
| Shaker | Thermo-Fisher | MaxQ 4450 | |
| Centrifuge | Eppendorf | 5702 | |
| Incubator | Thermo-Fisher | Forma Series II Water Jacketed CO2 Incubator Model 3100 |
|
| Trypsin EDTA, 1x (0.25%) | Corning | 25-053-CI | |
| T-75 Flasks | BD Biosciences | 353136 | |
| T-175 Flasks | BD Biosciences | 353112 | |
| Pipet tips | Rainin | P2, P10, P20, P200 and P1000 | |
| Pipet tips | Corning | Filtered tips P2, P10, P20, P200 and P1000 |
|
| Cell Counter | Invitrogen | Countess | |
| Countess Cell Counting Chamber Slides |
Invitrogen | C10313 | |
| Trypan Blue Stain (0.4%) | Gibco | 15250-061 | |
| Collagen Type I (Rat Tail) | BD Biosciences | 354236 | |
| 96 well plate, clear bottom, black | BD Biosciences | 353219 |
Etiketler
Bu Makaleden Alıntı Yapın
Modeling Ovarian Cancer Metastasis in Peritoneal Cavity Lining: A Method To Establish an In Vitro 3D Model of the Peritoneal Lining. J. Vis. Exp. (Pending Publication), e20375, doi: (2023).