Human Physiology By Dr C C Chatterjee ^NEW^
Mouse embryonic fibroblasts (MEFs) and Madin-Darby canine kidney (MDCK) epithelial cells were cultured in Dulbeccos modified Eagles medium (DMEM; ATCC, Manassas, VA) containing 10% fetal bovine serum (FBS; Sigma-Aldrich). Primary human dermal microvascular endothelial cells (HDMECs) were grown on gelatin-coated, fibronectin-coated, or gelatin-coated, collagen-coated dishes (BD Biosciences, Bedford, MA) in low-serum EGM2 medium (Lonza, Mount Royal, Canada). Primary human umbilical vein endothelial cells (HUVECs) were cultured in EBM-2 Medium (Lonza) with 1% FBS. Human embryonic kidney 293 cells and HeLa cells were grown in DMEM containing 10% FBS. All cells were cultured at 37 oC in a 5% CO2 environment and maintained in exponential growth.
human physiology by dr c c chatterjee
Knockout of ACE2 was obtained by infecting MDCK cells with lentivirus of shACE2 and a puromycin-selection marker. ACE2 expression was restored by lentivirus of pcDNA-ACE2. The mutants were generated by PCR-directed mutagenesis of pcDNA-ACE2. HEK293 cells were transfected with plasmid using PolyJet reagent (SignaGen Laboratories, Gaithersburg, MD). Plasmid encoding human ACE2 was a gift from S. Cohen (University of Pittsburgh). SMARTpool siRNAs against ACE2 were obtained from Dharmacon (Lafayette, CO). Human myosin light chain 2 was amplified from cDNA of adult rat heart using primers 5-CCTACACGCTCGCAGTGCTTACC-3 and 5-CTTTCCCTGATCCAGTTCCAGG-3, with the PCR product inserted into the BamHI and XhoI sites of pCMV-Tag2A to generate pCMV-MLC.
The overall purpose of the Human Physiology is to offer a popular and easily available textbook on human physiology, that will be easy to use, and will provide a sound foundation for the study of physiology with the minimum theoretical background. All chapters are tied together with appropriate introductory chapters, and are accessible to non-specialists as well as those with less background in science. The book is intended to be a second-year coursebook with relatively less coverage on cardiac, respiratory, and peripheral mechanisms. Key clinical areas have been covered especially in the last two chapters and the book includes several diagnostic tests, case reports and applications, which can be incorporated into classroom sessions.