After culturing for 44 h, cytochalasin
B was added in the cell culture medium.
Consisely, oocytes enucleation was performed under UV light, and a single fibroblast cell was injected into the perivitelline space of an enucleated oocyte in [M.sub.2] (Sigma, USA) medium supplemented with 10%FCS(HyClone, Beijing, China), 2 [micro]g/mL Hoechst 33342 (Sigma, USA) and 7.5 [micro]g/mL cytochalasin
B (Sigma, USA).
Various shock applications including heat / cold shock, hydrostatic pressure and chemical agents (Colchicine, Cytochalasin
B and [N.sub.2]O, etc.) have been used in the study in order to convert these haploid embryos, fertilized with spermatozoa and untreated, to diploid embryos [4,6-10].
The reconstructed embryos were equilibrated in PZM-3 supplemented with 5 mg/mL cytochalasin
B for 2 h at 38.5degC in humidified atmosphere of 5% CO2, 5% O2, and 90% N2 (APM-30D, ASTEC, Japan).
24 h after seeding (G1/S exposure), cultures were treated with different concentrations of BWD-1 (3, 15, and 75 [micro]g/[cm.sub.2]) 20 h prior to cytochalasin
B addition (Sigma, 6 [micro]g/ml) and the cultures were incubated for additional 28 hour (48 h treatment total).
D (cyto D) is a cell-permeable actin depolymerizing agent that inhibits endocytosis, but extended exposure to cyto D is cytotoxic.
To validate the results obtained by in Vitro Microflow kit, the analysis of MN in binucleated cells was performed by adding cytochalasin
B (Sigma-Aldrich; final concentration of 6 [micro]g/mL) after 44 h of culture .
Genotoxicity of HP is evaluated by cytochalasin
block micronuclei assay (CBMN) which was performed according to published procedures  with minor modifications.
1.25 mM MgS[O.sub.4] and 1.25 mM Ca[Cl.sub.2]) and either left untreated or stimulated with 1.74 [micro]M insulin, or with or without 20 [micro]M cytochalasin
B (inhibitor of glucose transporters).
Leibovitz's L-15 medium supplemented with 0.2% DMSO was used, either alone or supplemented with 0.2% cytochalasin
D (1 mg/mL in DMSO) to inhibit phagocytosis.
"Effects of cytochalasin
congeners, microtubule-directed agents, and doxorubicin alone or in combination against human ovarian carcinoma cell line in vitro." BMC Cancer.
Polyploid mosaics induced by cytochalasin
B in landlocked Atlantic salmon Salmo salar.