Cell counting kit 8 (cck8)

The in vitro cytotoxicity of nanofiber membranes was investigated through Cell Counting Kit-8 (CCK-8) assay (Dalian Meilun Biotechnology Co., Ltd., China) in L929 cell line. Sterilized nanofiber membranes were incubated with High-Glucose Dulbecco’s Minimum Essential Medium (H-DMEM, Hyclone, USA.) containing 10% fetal bovine serum (FBS, Gemini, USA) and 1% antibiotic (Solarbio, China) at 37°C for 2 days to prepare leaching solution of nanofiber membranes. L929 cells were seeded in 96-well plates with a density of 3 × 10³ cells/well and cultured at 37°C for 24 h. Then, L929 cells in 96-well plates were cultured with the leaching solution for another 24 h. The cell viability was measured by CCK-8 assay. Bone marrow stromal stem cells (MSCs) were isolated from SD rats and the 4th passage was used.39 (link) The cell proliferation of MSCs seeded on stereocomplexed nanofiber membranes was measured by CCK-8 assay. MSCs were seeded in 96-well plates with a density of 5 × 10³ cells/well and each well of the plate was covered by stereocomplex PLA/TTCP45. Low-Glucose Dulbecco’s Minimum Essential Medium (L-DMEM, Hyclone, USA) containing 10% FBS were used to culture MSCs. After cultured for 1, 3, and 5 days, live/dead staining and SEM images of MSCs cultured on stereocomplexed PLA/TTCP45 were observed by fluorescent microscopy and Hitachi SU3500 SEM (Hitachi, Japan).

The survival rate of A549 cells was assessed using the Cell Counting Kit 8 (Meilun Biotech, Dalian, China). The A549 cells were plated in 96-well plates at a density of 10×10³ cells/well and cultured for 24 hours. PHMG 0–32 μg/mL (High Polymer Bio, Shanghai, China) and T0901317 0~10 μM (Cayman chemical, Ann Arbor, Michigan, USA) were added to each well at different concentrations. After 6, 24 and 48 hours, the Cell Counting Kit 8 (CCK-8) solution (10 μL) was added to each well and cells were incubated at 37 °C for 1 hour. The absorbance at 450 nm was detected with a microplate reader (Tecan company, Austria).