Primary cultures of hemocytes from Mytilus galloprovincialis Lmk.: expression of IL-2Rα subunit

Effect of 17β-estradiol on adhesion of Mytilus galloprovincialis hemocytes to selected substrates. Role of alpha2 integrin subunit

“…ALS buffer doesn't effect the adhesion properties of hemocytes, since cell adhesion ability is determined enzymatically (Section 2.6). In order to isolate hemocytes, the cell suspension was centrifuged at 1200 rpm, for 15 min and the pellet containing the hemocytes was resuspended in Leibovitz L-15 medium supplemented with 350 mM NaCl, 7 mM KCl, 4 mM CaCl 2 , 8 mM MgSO 4 , 40 mM MgCl 2 , 10% FCS, 100 U/mL penicillin G, 100 mg/mL streptomycin, 40 mg/mL gentamycin, 0.1 mg/ ml amphotericin B, at pH 7 and 1000 mOsmol [30]. The medium was then filtered through 0.45 m filters and kept at 15 C. Cells were counted with the use of a Neubauer hemocytometer.…”

The role of signalling molecules on actin glutathionylation and protein carbonylation induced by cadmium in haemocytes of musselMytilus galloprovincialis(Lmk)

“…gentamycin, 0.1gml -1 amphotericin B at pH7 and osmolarity 1000mosmol l -1 (Cao et al, 2003). Cells were kept at 15°C for at least 3h before being used for the experiments.…”

Nitric oxide production by haemocytes from Mytilus galloprovincialis shows seasonal variations

“…The molluscs were taken to the laboratory within 1 h in tanks containing sterile seawater. Every 15 days throughout three consecutive years (1999e2001), mussels were collected and the haemolymph processed immediately on arrival at the laboratory [31]. The haemolymph was aspirated by a syringe from the adductor muscle and mixed with 1 ml of ALS (Alseve) buffer (20.8 g/l glucose, 8 g/l sodium citrate, 3.36 g/l EDTA and 22.5 g/l NaCl, pH 7.0 and 1000 mOsmol).…”

“…The suspension containing the haemocytes was centrifuged (450 Â g for 10 min) at room temperature. The pellet containing the haemocytes was re-suspended and cultured for 3 days at 20 C in Leibovitz L-15 medium supplemented with 20.2 g/l NaCl, 0.54 g/l KCl, 0.6 g/l CaCl 2 , 1 g/l MgSO 4 , 3.9 g/l MgCl 2 , 20.8 g/l glucose, 10% foetal calf serum (FCS), 100 U/ml penicillin G, 100 mg/ml streptomycin, 40 mg/ml gentamicin and 0.1 mg/ml amphotericin B at pH 7.0 and 1000 mOsm [31]. The cell viability was assayed by the Trypan blue exclusion technique.…”

“…After 3 days in culture [31], the cells were re-counted and then distributed into 1 ml aliquots of FCS-supplemented ALS medium (culture medium) containing ALS buffer and 17.4 g/l arginine, 29.22 mg/ml glutamine, 10% FCS, 100 U/ml penicillin, 100 mg/ml streptomycin, 40 mg/ml gentamicin and 0.1 mg/ml amphotericin B, containing 10 6 cells. These suspensions were added to the necessary amount of each stimulator and put back into the incubator at 20 C for the time required in each experiment.…”