2D Co-culture of Mouse Endothelial and Osteocytic Cells
The mouse endothelial cell line bEnd.3 was obtained from ATCC, Catalog number CRL-2299. MLO-Y4 was a gift from Dr Lynda F Bonewald. bEnd.3 cells were maintained in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (Gibco Cat. 10099141) and 1% penicillin/streptomycin (P/S). MLO-Y4 cells were maintained in alpha–minimal essential medium (alphaMEM) containing 10% FBS and 1% P/S. For 2D coculture, bEnd.3 cells were seeded on coverslips in 24-well plates. After 24 hours, MLO-Y4 cells were seeded in the wells that cultured bEnd.3 cells. The cells were maintained at 37 °C in a 5% CO2 humidified incubator.
Liao P., Chen L., Zhou H., Mei J., Chen Z., Wang B., Feng J.Q., Li G., Tong S., Zhou J., Zhu S., Qian Y., Zong Y., Zou W., Li H., Zhang W., Yao M., Ma Y., Ding P., Pang Y., Gao C., Mei J., Zhang S., Zhang C., Liu D., Zheng M, & Gao J. (2024). Osteocyte mitochondria regulate angiogenesis of transcortical vessels. Nature Communications, 15, 2529.
Publication 2024
Corresponding Organization :
Other organizations :
Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Second Affiliated Hospital of Zhejiang University, University of Western Australia, Shanxi Medical University, Zhejiang Chinese Medical University
Cell growth and/or cell behavior in the 2D coculture system
control variables
Cell culture conditions (temperature, CO2 levels, and humidity)
Culture media (high-glucose DMEM for bEnd.3 cells, alpha-MEM for MLO-Y4 cells)
Serum (10% FBS) and antibiotics (1% P/S) in the culture media
controls
Positive control: Not specified
Negative control: Not specified
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