Mesenchymal Cells and Fibrocytes in Peripheral Blood. Culture Conditions

ObjectivesThe fibrocytes, the mesenchymal cells, and the colony forming units are included in the concept fibroblast-like, peripheral-blood circulating stem cells. The aim of this work was to establish the culture conditions.Material and methodPeripheral blood mononuclear cells were isolated from 25 samples collected from the cephalic vein. These were cultured in McCoy's 5A enriched with 20% of fetal bovine serum. The culture was read at 7, 14, and 21 days, evaluating the proliferation, confluence, and the round cell/spindle cell ratio. Moreover the 21st day surface markers and phagocytic activity testing was carried out.ResultsGrowth was achieved in all the cases but only in 2 dishes were confluence areas retrieved. Spindle cells were 36 (3.71%) (mean [SD]). All the cells showed intense signal against vimentin, CD34 marker, and the NBT test were positives.ConclusionsClear CD34 expression, positivity against a connective tissue marker such as vimentin and positivity in a phagocytic assay like the NBT-test, support that stem cells, monocytes, and fibroblasts share common characteristics.

ObjetivosEl término fibroblast-like, célula madre de sangre periférica, engloba a las unidades formadoras de colonias fibroblásticas, los fibrocitos y las células mesenquimales. El diseño de este trabajo intenta establecer las condiciones de cultivo.Material y métodoEn 25 muestras de sangre venosa se aislaron células mononucleares y se sembraron en medio de cultivo McCoy's 5A con suero bovino fetal al 20%. A los 7, 14 y 21 días se evaluó la proliferación, la confluencia y la relación células redondas/células alargadas. El día 21 se valoró la morfología, los marcadores de superficie y la actividad fagocítica.ResultadosEn todos los cultivos se detectó crecimiento celular positivo para la vimentina, CD34 y NBT. La media ± desviación estándar del 36 ± 3,71% eran células alargadas (fibrocitos) y sólo en 2 casos se registraron áreas de confluencia.ConclusionesLos métodos de cultivo descritos son adecuados para los seudofibroblastos de sangre periférica. La positividad de CD34, prueba NBT y vimentina demuestra que los seudofibroblastos tienen características comunes con la célula madre, los monocitos-macrófagos y los fibroblastos.