Phenotype and ultrastructure of stem cells derived from amniotic fluid of Nitra rabbit

Abstract

The isolation of amniotic fluid-derived mesenchymal stem cells (AF-MSCs) has been already shown in human and several other species including rabbit. However, prior to the preclinical research on various animal models it is desirable to define AF-MSCs by a panel of surface protein markers. Therefore, the aim of this preliminary study was to detect the expression of several protein markers on the surface of AF-MSCs isolated from local breed of Nitra rabbit. Amniotic fluid was collected from humanely sacrificed rabbits (n = 3) and AF-MSCs were cultured to a third passage. Flow cytometry was used to detect surface protein marker expression and for viability testing. Rabbit AF-MSCs (rAF-MSCs) were also analyzed by transmission electron microscopy to define the ultrastructure. rAF-MSCs showed both sufficient viability (more than 80%) and low apoptosis rates at third passage and highly expressed CD29 (88.17 ± 7.17%) and CD44 (80.00 ± 2.28%). However, a dim expression of CD90 (17.24 ± 1.31%) and negative expression of CD73 (1.21 ± 0.56% and 4.41 ± 1.46%), CD105 (1.67 ± 0.37%) and CD166 (0.96 ± 2.26%) was observed. Additionally, ultrastructure analysis revealed eccentrically located nucleoli, an abundance of thin pseudopodia on cells’ surfaces and proved the presence of typical mesenchymal stem cell features. In conclusion, this set of data contributes to more detailed information on rAF-MSCs, which were previously proposed feasible for preclinical stem cell research and as a suitable source for the cryopreservation of animal genetic resources in gene bank.