Bioflex amino culture plates

Numerous routine treatment methods were tested in the laboratory, such as soaking in hydrochloric acid (plate) for 24 h, followed by soaking in water for 24 h, then drying, gelatin coating or cleaning. Moreover, drying after soaking in 75% alcohol for 24 h, then gelatin coating was performed. However, when using these methods cells did not adherent to the wells and grew poorly. Later, the plate was washed and dried. Then, the gelatin was changed into rat tail collagen for coating, it was found that the cells could adhere to the wall and grow efficiently. The BioFlex amino-culture plates (Flexcell International Corporation) were coated with 2 mg/ml rat tail collagen solution (Roche Diagnostics). Then, 20 µl of collagen solution was added to each well and smeared evenly with an aseptic cell scraper. Subsequently, the plates were blow-dried on a clean bench and irradiated with ultraviolet light for 1 h.

Activated HSCs were plated onto collagen-coated BioFlex amino-culture plates (Flexcell International, McKeesport, PA, USA) and grown in DMEM without FBS for 24 h. HSCs were then subjected to a 10% tensile strain, 0.5 Hz for 24 h by using Flexercell FX-2000 system (Flexcell International), as previously described (18 (link)).
Total RNA was extracted from HSCs with or without tensile strain treatment by using miRVana™ RNA Isolation kit (Thermo Fisher Scientific, Inc., Rockford, IL, USA) following the manufacturer's instructions. The extracted RNA was then quantified by using NanoDrop spectrophotometer (NanoDrop Technologies: Thermo Fisher Scientific, Inc., Wilmington, DE, USA) at 260 nm.