Penicillin streptomycin 100x

Polycaprolactone (PCL, M_w = 80000),
dichloromethane (DCM), N,N-dimethylformamide
(DMF), methanol, and chloroform
were purchased from Sigma-Aldrich (St. Louis). Glass syringes (with
an inner diameter of 14.6 mm), three-layers coaxial needle, and coaxial
kit were acquired from Linari NanoTech (Pisa, Italy). The KDS-100-CE
syringe pump was purchased from KD Scientific (Holliston, MA). Rifampicin
was acquired from EMD Millipore Corp. The D-ES30PN-20W potential generator
was purchased from Gamma High Voltage Research Inc. (Ormond Beach,
FL). Recombinant Trypsin–EDTA 1X, penicillin/streptomycin 100X, l-glutamine 100X, fetal bovine serum (FBS), and Dulbecco’s
modified Eagle’s medium (DMEM) were purchased from Euroclone
(Milan, Italy).

Three human breast cancer cell lines, MDA-MB231, MDA-MB468 and MCF-7, all derived from adenocarcinoma metastasis and on normal human breast epithelial cells MCF-10A were used. In particular, MCF-7 and MCF-10A cells were expanded at 37 °C in a humidified atmosphere of 5% CO₂ in culture medium DMEM (Dulbecco’s Modified Eagle’s Medium, Lonza), whereas MDA-MB-231 and MDA-MB-468 in RPMI 1640 (Lonza, Munchensteinerstrasse, Basel, Switzerland), supplemented with fetal bovine serum (FBS) (Invitrogen, Camarillo, CA, USA) at 10%, Penicillin/Streptomycin 100x (Euroclone, Devon, UK), Glutamax 100x (Invitrogen) non-essential amino acids 100x (Invitrogen). Moreover, in the case of MCF-10A the DMEM was supplemented also with human insulin 10 μg/mL (Life Technologies Corporation, Carlsbad, CA, USA), human epidermal growth factor 20 ng/mL (Life Technologies), and hydrocortisone 0.5 μg/mL (Sigma-Aldrich, St. Louis, MO, USA) according to the procedure reported in Rothwell et al. (2014), while for MDA-MB468 the medium was implanted with Ham’s F-12 medium (1:1 mixture). Phosphate buffer (PBS Phosphate buffered saline Ca²⁺ and Mg²⁺ free) and trypsin (Ca²⁺ and Mg²⁺ free) were supplied by Euroclone. Finally, the cells were kept in an incubator at a humidified atmosphere of 95% air and 5% CO₂ at 37 °C.

Lymphoma cell lines were cultured according to the recommended conditions, as previously reported [14 (link), 15 (link)]. All media were supplemented with fetal bovine serum (10%), Penicillin‐Streptomycin 100X (Euroclone, ECB3001D). Cell line's identity was confirmed by short tandem repeat (STR) DNA fingerprinting using the Promega GenePrint 10 System kit (B9510) [9 (link)], and all the experiments were performed within 1 month from being thawed. Cells were periodically tested to confirm mycoplasma negativity using the MycoAlert Mycoplasma Detection Kit (Lonza, Visp, Switzerland).