Different RNA and protein expression of surface markers in rabbit amniotic fluid‐derived mesenchymal stem cells

Abstract

Over the years, there has been much confusion in defining molecular markers of mesenchymal stem cells (MSCs) for other than human species due to a lack of species-specific antibodies. Therefore, the aim of our study was to define rabbit amniotic fluid-derived mesenchymal stem cells (rAF-MSCs) and to reflect upon the current identification of AF-MSCs by providing a summary of detected surface markers in different species. The expression of rAF-MSC surface markers was analyzed at the protein and mRNA level. Flow cytometry analyses showed that rAF-MSCs were positive for CD29 and CD44, low positive for CD90, but negative for CD73, CD105, and CD166. Interestingly, RT-PCR (reverse transcription-polymerase chain reaction) exposed a discprepancy between transcribed mRNA and protein expression, as the cells expressed mRNA of all MSC markers: CD29, CD44, CD73, CD90, CD105, and CD166. Our results also confirmed the mesenchymal nature of isolated cells by morphology, ultrastructure, and intracellular marker expression profile. In addition, the expression of few pluripotency markers was also detected. We also found that passaging did not affect apoptosis and viability. Similarly, changes in karyotype were not observed during passaging. In conclusion, the provided characteristics may be used as a comprehensive set of criteria to define and characterize rAF-MSCs, required for the identification of these cells in preclinical investigations. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1601-1613, 2017.

Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources

“…Clinically healthy and young (3-to 8-month-old) rabbits (n = 20) of the New Zealand White (NZW) line reared as described previously [45] were used in this study. The treatment of the animals was approved by the Ministry of Agriculture and Rural Development of the Slovak Republic no.…”

“…The expression of selected markers was also analyzed in the third passage of HUVECs. Staining of cells with the primary antibodies was performed as described previously [45]. The proper secondary antibodies were used for the purified primary antibodies: rat anti-mouse IgG1-FITC (clone M1-14D12; eBioscience, Wien, Austria), goat anti-rabbit IgG-FITC (polyclonal; 405002, Bio-Rad, Hercules, CA, USA) and goat anti-mouse IgG-FITC (polyclonal; STAR117F, Bio-Rad, Hercules, CA, USA).…”

Secretome Analysis of Rabbit and Human Mesenchymal Stem and Endothelial Progenitor Cells: A Comparative Study

“…These features make them both interesting for stem cell therapy, tissue engineering, and regenerative medicine. For such studies, rabbit stem and progenitor cells became also a favorable choice [2,4,5,12].…”

“…For almost the last two decades, researchers have focused on the study of mesenchymal stem cells (MSCs) due to their specific biological features. This specific cell type can be presently obtained from various human as well as rabbit biological sources, most often from bone marrow, amniotic fluid, or others [1][2][3][4]. However, recently, adipose tissue became preferred for the isolation of MSCs as a more feasible biological source from human or rabbit [1,5].…”

Protection and Sustainability of Slovak Animal Genetic Resources in Order to Ensure the Self-Sufficiency in Quality Food in Slovakia

“…In our institute NPPC-RIAP Nitra we are dealing with cryopreservation and storage of the genetic material like semen from bulls Makarevic et al, 2018;Olexikova et al, 2016;Spalekova et al, 2012); rabbits (Kulikova et al, 2014;, rams (Kulikova et al, 2018;Makarevic et al, 2012) , chicken (Svoradova et al, 2018) or embryos from cattle (Makarevic et al, 2014), rabbit (Chrenek et al, 2014), stem cells from rabbits (Kovac et al, 2017;Vasicek et al, 2011) and DNA from honey bee (Bauer et al, 2017).…”