Mda mb 231 gfp cell line

Three different human epithelial cell lines were investigated. (i) MCF10A cells (non-tumorigenic epithelial cell line derived from human mammary gland ATCC CRL®-10317™) were cultured in 1:1 DMEM:F12 medium (Gibco) supplemented with 5% horse serum (Invitrogen), 2 mM L-Glutamine (Sigma Aldrich), 20 ng/ml EGF (Sigma Aldrich), 0.5 mg/ml hydrocortisone (Sigma Aldrich), 100 ng/ml cholera toxin (Sigma Aldrich), 10 μg/ml insulin (Sigma Aldrich), 1% PenStrep (100 U/ml of penicillin, 100 μg/ml streptomycin, Gibco). (ii) MCF7 cells (derived from primary tumor, human invasive breast ductal carcinoma ATCC® HTB-22™) were grown in DMEM medium (Gibco) supplemented 10% FBS (Gibco) + 2mM L-Glutamine (Sigma Aldrich), 1% PenStrep. (iii) MDA-MB-231 (human breast adenocarcinoma cell line derived from a metastatic site ATCC® HTB-26™) were cultured in RPMI medium supplemented 10% FBS (Gibco) + 2mM L-Glutamine (Sigma Aldrich), 1% PenStrep. MDA-MB-231/GFP Cell line (CELL BIOLABS, INC.) were grown in DMEM (high glucose), 10% FBS, (Sigma Aldrich), 2 mM L-Glutamine, 1% Non Essential Amino Acids, 1% PenStrep. All cell lines were cultured at 37 °C and 5% CO₂ and split every three days 1:5 by using trypsin/EDTA (Sigma Aldrich).

The cell lines chosen for NPs–cell interaction were MDA-MB-231 (CD44^+ve/high), BT-474 (CD44^−ve/low) and NIH 3T3 (CD44^−ve). The MDA-MB-231-GFP cell line used for FACS experiments was procured from Cell Biolabs, Inc., CA, USA (catalogue no. GFPAKR210). MDA-MB-231 and BT-474 cancer cell lines were generous gifts from Prof. Annapoorni Rangarajan, Indian Institute of Science, Bangalore, India. NIH 3T3 cell line was a kind of gift from Dr Arka Shubra Ghosh, GROW labs, Narayana Netralaya, Narayana Health City, Bangalore, India. The basal medium used for all cell culture experiments was Dulbecco’s Modified Eagle’s medium supplemented with 10% Fetal Bovine Serum, and antibiotics streptomycin sulfate and benzyl at final concentrations of 100 µg/ml and 100 U/ml respectively. Cell cultures were maintained at 37 °C at 5% CO₂ throughout all experiments. The cells were passaged using 0.25% Trypsin-EDTA at around 80% confluency. All the cells used in this study were tested for mycoplasma contamination by 4, 6-diamidino-2-phenylindole (DAPI) staining, and were found to be negative.