Minimal essential medium eagle mem

Animal management was performed according to the ethical guidelines of our institutions, and animal use was performed according to the Mexican norm NOM-062-ZOO-1999, and its technical specifications for the production, care and use of laboratory animals. The F. hepatica reference strain was maintained as an anthelmintic bioassay reference at the National Center for Disciplinary Research in Veterinary Parasitology (INIFAP, Jiutepec, Morelos, Mexico) and previously registered as NCBI-BioSample SAMN16822856 [3 (link)], 90 metacercariae were orally inoculated in a six-months-old rabbit. Parasitic development was confirmed four months later by the presence of liver fluke eggs in the feces. The rabbit was sacrificed, and the liver was removed and dissected. The parasites were recovered from the liver, and a group of twenty parasites was collected in 20 mL of Minimal Essential Medium Eagle (MEM, Sigma Chemicals, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS GIBCO ).

We established primary human fibroblasts from healthy volunteers and patients at Kyushu University Hospital, and also purchased human fibroblasts from the cellular bank of the Coriell Institute (Camden, NJ, USA) (Supplementary Table 1). Informed consent was obtained from all the healthy volunteers and patients before donating skin fibroblasts. To establish the fibroblast cell culture, the epidermis and subcutaneous adipose tissues were removed from biopsied skin tissue, and the remaining dermis tissue was placed on a clean culture dish. The dermis tissues were radially impressed the surgical knife to stick them to the bottom of the culture dish. Fibroblasts were maintained in Fibroblast Growth Medium (FGM) that contained 15% fetal bovine serum (FBS) (Japan Bioserum, Hiroshima, Japan), 0.1 mM MEM Non-Essential Amino Acids (NEAA) (Thermo Fisher Scientific, Waltham, MA, USA), and 1% Pen Strep (Thermo Fisher Scientific) in Minimal Essential Medium Eagle (MEM) (Sigma Chemical Co., St. Louis, MO, USA).

The toxicity of the rippled samples with AuNWs was tested in primary human lung fibroblast (MRC-5, American Tissue Culture Collection, Manassas, VA, USA). Human primary fibroblasts were incubated in Minimal Essential Medium Eagle (MEM, Sigma-Aldrich, St. Louis, MO, USA) supplemented with 2 mM L-glutamine (a stable dipeptide, sourced from Sigma-Aldrich, St. Louis, MO, USA) under standard conditions (37 °C, 5% CO₂).
Samples (PEN 22.5° and Au/PEN 22.5°) with human health cells were prepared as described by the authors of [26 (link)]. In this case, cell viability was measured using a resazurin assay [43 ]. The medium was removed, then the samples were washed with PBS and incubated with a resazurin solution (final concentration 25 μg/mL) in a medium without phenol red (MEM, Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and incubated for 4 h. Thereafter, fluorescence was measured using the Fluoroskan Ascent (Thermo Labsystems, Waltham, MA, USA) and excitation and emission wavelengths of 560 and 590 nm, respectively. Cells that were grown on standard tissue culture polystyrene (TCPS) were used as control. All samples were prepared in triplicate. Cell viability was represented as a percentage of metabolic activity of control cells. Mean values and standard deviations were calculated.