Fgm cd bullet kit

To determine the in vitro and in vivo effects of hsa-miR-520d-5p expression and to explore its safety for future systemic administration, we used three cell lines and lentiviral vectors. Human iPSCs (hiPSCs) (HPS0002) were provided by the RIKEN BioResource Center Cell Bank (Ibaraki, Japan), and both human umbilical vein endothelial cells (HUVECs) and normal human dermal fibroblast (NHDF) cells were provided by TAKARA BIO Inc. (Tokyo, Japan). To examine the effect of miR-520d-5p on normal cells in vitro and in vivo, we used a human fibroblast cell line (NHDF-Ad derived from adults) and HUVECs, cultured in fibroblast basal medium (FBM)-2 medium using the fibroblast growth medium chemically defined (FGM-CD) Bullet Kit and EBM-2 using EBM-2 SingleQuots, respectively (TAKARA BIO Inc.). The hiPSCs were cultured in ReproStem medium (ReproCell, Tokyo, Japan) with 5 ng/mL basic fibroblast growth factor (bFGF)-2. In addition, the human mesangial cell line 293FT (Invitrogen Japan K.K., Tokyo, Japan) was used for producing miR-520d-expressing lentivirus as previously reported [13 (link)]. The 293FT cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 % fetal bovine serum (FBS), 0.1 mM minimum essential medium (MEM) non-essential amino acids solution, 2 mM l-glutamine, and 1 % penicillin/streptomycin.

To determine the in vitro effects of hsa-miR-520d-5p on UV-irradiated fibroblasts and to explore its protective application for anti-aging therapy, we used three cell lines and a lentiviral vector. NHDF cells were provided by TAKARA BIO (Tokyo, Japan). To examine the effect of hsa-miR-520d-5p on normal cells in vitro, we used NHDF cells cultured in fibroblast basal medium (FBM)-2 using the fibroblast growth medium chemically defined (FGM-CD) Bullet Kit (TAKARA BIO). In addition, the human mesangial cell line 293FT (Invitrogen Japan K.K., Tokyo, Japan) was used to produce the hsa-miR-520d-expressing lentivirus. The 293FT cells were cultured in Dulbecco’s modified Eagle’s medium (WAKO, Tokyo, Japan) supplemented with 10% fetal bovine serum, 0.1 mmol/l minimum essential medium (MEM) non-essential amino acids, 2 mmol/l l-glutamine, and 1% penicillin/streptomycin.