Características de células fibroblast-like derivadas do cultivo do saco vitelino em diferentes períodos gestacionais

Abstract

Based on the knowledge regarding the erythropoietic activity of the vitelline membrane, which is considered the first hematopoietic and vascular site that appears in mammals during organogenesis, added to the erythropoiesis support function performed by mesenchymal stem cells, the present work aims to verify the cell expansion of the yolk sac using the explant technique, in view of the cell niches found in the fetal annex. For the cultivation of the yolk sac, 5 bitches with gestational ages of 30, 40 and 60 days were used, and were checked by ultrasound and Crow - Rump of each fetus according to Evans & Sack (1979). The bitches were submitted to ovariosalpingohisterectomy to collect the fetal attachment. All material was collected at the Faculty of Veterinary Medicine and Animal Science at the University of São Paulo - USP. The yolk sac was fragmented manually (mechanical fragmentation), with the aid of a scalpel blade until obtaining a homogenate of tissue fragments and cell groups, which were implanted in the 5 ml culture bottle with the modified Eagle Dulbeco medium - DMEM High with 10% fetal serum, streptomycin and penicillin. The cells were incubated at 38.5 degrees Celsius, at 100% humidity and 5% CO 2. The morphological description of the cell culture was performed periodically using the Nikon TS100 phase contrast microscope. The cells maintained in these conditions grew in the form of monolayer, with different fusiform cell types and large cells with cytoplasmic extensions like fibroblast-like.