Mrc 5 ccl 171

Human ARPE-19 (CRL-2302) retinal pigmented epithelial cells, MRC-5 (CCL-171) embryonic lung fibroblasts cells were obtained from American Type Culture Collection (ATCC) and were maintained according to the supplier’s recommendations^{63 (link)}. AD169 (VR-538) was obtained from ATCC and propagated in MRC-5 cells. AD169r-GFP virus was a generous gift of Thomas Shenk of Princeton University, and propagated in ARPE-19 cells. The live-attenuated HCMV virus vaccine, V160, was generated by restoring the wild-type pentamer in the AD169 strain as reported^{27 (link)}. Viruses were produced in MRC-5 or ARPE-19 cells, and viral particles were harvested and purified by centrifugation at 55,000 × g for 90 min through 20% sorbitol cushion in 50 mM tris (pH 7.2) and 1.0 mM MgCl₂. The pellet was resuspended and stored at −80 °C.

The human OC cell line (SKOV3 (HTB-77)) and human lung normal fibroblast cell line (MRC-5 (CCL-171)) were obtained from the American Type Culture Collection (ATCC). We also used the second OC cell line derived from humans (PEO1 (10032308)) purchased in the European Collection of Authenticated Cell Cultures (ECACC). The SKOV3 cell line was a hypodiploid OC derived from a 64-year-old Caucasian female with an ovarian serous cyst adenocarcinoma. The number of chromosomes was 43, occurring in over 60% of the cells. These adenocarcinoma cells were positive for many antigens generally used to identify epithelial cancer—for example: EMA (epithelial membrane antigen), VIM (vimentin) and cytokeratin alike.
The second OC cell line used in the experiments was PEO1 derived from a malignant effusion from the peritoneal ascites of a patient with a poorly differentiated serous adenocarcinoma after treatment with chlorambucil, 5-fluorouracil and cisplatin. This cell line was positive for hormone receptors like the estrogen receptor.
Normal human fibroblasts were used as control cells. The MRC-5 fibroblast line was derived from normal lung tissue of a 14-week-old male fetus. This is a normal diploid human cell line with 46 XY karyotypes.

A549 (CCL-185), MCF-7 (HTB-22), 4T1 (CRL-2539), HeLa (CCL-2), and MRC-5 (CCL-171) cells were purchased from American Type Culture Collection (ATCC). A2780 and A2780cisR cells (originally purchased from ATCC, HTB-174) were provided by W. H. Ang (Department of Chemistry, National University of Singapore). A549cisR cells were cultured as previously reported (16 , 17 (link), 58 (link)). Human breast carcinoma MCF-7 cells, human cervix carcinoma HeLa cells, human lung carcinoma A549 cells, and cisplatin-resistant A549cisR cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% FBS and penicillin/streptomycin (100 IU ml⁻¹). Human ovarian carcinoma A2780 and A2780cisR cells were cultured in RPMI 1640 containing 10% FBS, 2 mM l-glutamine, and penicillin/streptomycin (100 IU ml⁻¹). Human lung fibroblast MRC-5 cells were cultured in MEM containing 10% FBS, 1% non-essential amino acids (NEAA), 1% l-glutamine, 1% NaPyr, and penicillin/streptomycin (100 IU ml⁻¹). In every two passages of A2780cisR and A549cisR cells, 5 μM cisplatin was added to maintain the platinum resistance. All the cells were cultured in humidified incubators at 37°C with 5% CO₂. Cell counting was performed with a hemocytometer.