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.
Koncurat
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Departamento de Microbiología e Immunología, Facultad de Ciencias Exactas, Fisico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, Argentina.
Greco
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CR
Vivas
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D000328:Adult; D017077:Culture Media, Conditioned; D046508:Culture Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D008213:Lymphocyte Activation; D010920:Placenta / Q000737:chemistry*; D011247:Pregnancy; D014020:Tissue Extracts
D017077:Culture Media, Conditioned; D014020:Tissue Extracts