Human umbilical vein endothelial cells (HUVEC, ATCC CRL-1730) were grown in DMEM (Lonza) with FBS (10%, PAA), ultraglutamine (1%, Lonza) and gentamicin sulfate (Lonza) at 37 °C and CO2 (5%). Murine alveolar MH-S macrophages (ATCC CRL-2019) were grown in RPMI 1640 (Lonza) supplemented with heat-inactivated FBS (10% FBS), sodium-pyruvate (1 mM, Lonza), ultraglutamine (1%) and gentamicin sulfate (50 mg mL−1) at 37 °C and CO2 (5%). THP-1 cells (DSMZ, ATCC 16) were maintained in RPMI 1640 supplemented with FBS (10%), ultraglutamin (2 mM) and gentamicine sulfate at 37 °C and CO2 (5%). U-937 cells (ATCC CRL-1593.2) were cultered in RPMI 1640 supplemented with FBS (10%) and gentamicin sulfate at 37 °C and CO2 (5%).
Cells were authenticated and tested for mycoplasma contamination by ATCC, passaged every second day until passage 30. Normal human serum (NHS) was prepared from FHR1/3 sufficient as well as FHR1/3 deficient (ΔFHR1/3 NHS) blood derived from healthy volunteers, as determined by PCR52 (link) and Western blot analyses. After coagulation blood was centrifuged (10 min, 2000×g, 4 °C), and NHS kept frozen in aliquots at −80 °C. C. albicans cph1Δ/efg1Δ53 (link), 54 (link), which cannot form hyphae, was grown overnight in YPD medium (D glucose (2%), peptone (1%), yeast extract in H2O (5%) at room temperature.
Cells were authenticated and tested for mycoplasma contamination by ATCC, passaged every second day until passage 30. Normal human serum (NHS) was prepared from FHR1/3 sufficient as well as FHR1/3 deficient (ΔFHR1/3 NHS) blood derived from healthy volunteers, as determined by PCR52 (link) and Western blot analyses. After coagulation blood was centrifuged (10 min, 2000×g, 4 °C), and NHS kept frozen in aliquots at −80 °C. C. albicans cph1Δ/efg1Δ53 (link), 54 (link), which cannot form hyphae, was grown overnight in YPD medium (D glucose (2%), peptone (1%), yeast extract in H2O (5%) at room temperature.
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