Human placenta conditioned medium as a source of lymphocyte blastogenesis in vitro

The purpose of this work was to obtain placental extracts with the capacity of producing lymphocyte blastogenesis. Placental homogenates were incubated at 37 degrees C in McCoy's 5A supplemented with 5% fetal bovine serum and 25 mM Hepes. After a 6-day incubation period, the cultures were harvested and the sterility and cellular viability were controlled. The supernatant was centrifuged at 22,000g at 4 degrees C for 30 minutes. Sterilization was achieved by filtration (0.22 microns). This medium was denominated human placenta conditioned medium (HPCM-1). The other half of the original placenta was processed by the Burgess method and the extracts thus obtained were denominated HPCM-2. The activity of the extracts thus obtained was tested in peripheral lymphocyte cultures from women after the third pregnancy; 5, 10 and 20% HPCM-1 or HPCM-2 was added to these cultures and the index of lymphocyte blast transformation (ILBT) was determined. The highest stimulation occurred at 10% HPCM. The ILBT obtained with HPCM-1 at 10% was more constant than that obtained with HPCM-2 at the same concentration. In conclusion, it can be assumed that this methodology is appropriate to obtain conditioned media from human placenta with good blastogenic activity on peripheral lymphocytes in vitro.