THE EFFICIENCY OF IMMUNOMAGNETIC SORTING OF RABBIT BONE MARROW CELLS FOR THE ESTABLISHMENT OF MESENCHYMAL STEM CELL CULTURE

Abstract

The aim of this study was to assess if immunomagnetic sorting of SSEA-4 or MSCA-1 positive bone marrow cells could facilitate and expedite the establishment of rabbit MSCs culture. Briefly, rabbit bone marrow cells were enriched for SSEA-4+ and MSCA-1+ cells using two clones of SSEA-4 antibodies (REA101 and MC-813-70) and MSCA-1 antibody (clone W8B2) via magnetic-activated cell sorting. Flow-cytometric analysis revealed a high level of sorting efficiency, since the SSEA-4+ cells were significantly (P<0.001) enriched from 17 to 95% and MSCA-1+ cells from 8 to 74%. According to morphological changes, both sorted fractions (positive and negative) successfully initiate rabbit MSCs culture, but not expedite the proliferation rate as compared to standard MSCs culture initiated from unsorted cells. Moreover, all samples (sorted and unsorted) expressed high levels of the typical MSCs markers (CD29 and CD44) and low expression of hematopoietic marker CD45. In conclusion, although this study revealed the possibility to enrich SSEA-4+ and MSCA-1+ positive cells from the rabbit bone marrow, the standard method of MSCs isolation using the plastic adherent ability of the unsorted bone marrow cells seems to be still useful and less cost effective.