2 mM L-glutamine (Sigma) was added to the monocyte cell culture.

LPS (1microgram/ml, Sigma, St. Louis, MO) (along with R848, Curdlan, yeast RNA and yeast cells in different conditions of culture) was used to induce DC activation.

These dendritic cells were obtained from PBMC from which monocytes had been isolated that differentiated into DC after cell culture.

Differentiation of monocytes into dendritic cells is promoted, along with granulocyte-macrophage colony-stimulating factor, by addition of recombinant IL-4 (1000U/ml, R&D Systems).

Differentiation of monocytes into dendritic cells is promoted, along with recombinant IL-4, by addition of granulocyte-macrophage colony-stimulating factor (GMCSF 1000U/ml, Chemicon).

Monocytes were cultured in RPMI 1640 medium (GIBCO BRL) supplemented with 2 mM L-glutamine (Sigma), 1% (vol/vol) non-essential amino acids, 100 mM sodium pyruvate, 50 U/ml of penicillin and 50 mg/ml of streptomycin (Gibco BRL) containing 10% (vol/vol) FCS (Hyclone).

50 mg/ml of streptomycin (Gibco BRL) containing 10% (vol/vol) FCS (Hyclone) were added to the monocyte cell culture.

50 U/ml of penicillin were added to the monocyte cell culture.

100 mM sodium pyruvate were added to the monocyte cell culture.

1% (vol/vol) non-essential amino acids were added to the monocyte cell culture.

These monocytes were obtained after elutriation of a leukapheresis product (patients with stage III or stage IV melanoma) and are used to obtain dendritic cells from.

Monocytes were isolated from PBMC to generate dendritic cells.

PBMC were isolated from buffy coat. Subsequently, monocytes were isolated and differentiated into dendritic cells.

PBMC were isolated from this buffy coat. Subsequently, monocytes were isolated from the PBMC and dendritic cells generated from the monocytes.

The patients were vaccinated with a vaccine composed by autologous DCs pulsed with apoptotic allogenic prostate carcinoma cell line LNCap.

These cells were used to produce a vaccine for prostate carcinoma and were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap cells.

A portion of the DCs is pulsed with irradiated tumor cells (apoptotic bodies) prepared from the removed lesion and matured for 48 hours in presence of TNF-alpha.

TNF-alpha is used during maturation of the DCs.

These cells are prepared from the removed lesion from patients with stage III or stage IV melanoma and are subsequently used to pulse a portion of the immature dendritic cells.

These DCs have been generated from monocytes after elutriation and have not been pulsed with irradiated tumor cells (apoptotic bodies).