Long bone mesenchymal stem cells (Lb-MSCs): clinically reliable cells for osteo-diseases Shirin Toosi . Hojjat Naderi-Meshkin . Fatemeh Kalalinia . Mohammad Taghi Pievandi . Hossein Hosseinkhani . Ahmad Reza Bahrami . Asieh Heirani-Tabasi . Mahdi Mirahmadi . Javad Behravan Received: 25 December 2016 / Accepted: 8 August 2017 Ó Springer Science+Business Media B.V. 2017 Abstract Mesenchymal stem cells (MSCs) have been designated as the most reliable cells in clinics to treat osteo-diseases because of their versatile nature. MSCs, isolated from long bone (Lb-MSCs) are rarely reported and named as RIA-MSCs because of the reamer–irrigator–aspirator (RIA) device. The poten- tial of these cells in the treatment of non-union bone fractures made them the ideal candidates to be studied for clinical practices. In this work, effect of cryop- reservation on the proliferation and differentiation capabilities of long bone MSCs (Lb-MSCs) has been studied. For this purpose, Lb-MSCs were isolated via RIA device and characterized using flow cytometry and differentiation assays. Cells were cryopreserved for 3, 6 and 12 months and thereafter were character- ized using differentiation assays and genetic markers specific for osteogenic, chondrogenic, and adipogenic potential quantitatively by qRT-PCR. Lb-MSCs were found expressing MSC characteristic markers defining their identity. The population doubling time (PDT) was about 2.5 ± 0.5 days and colonies appeared after 7–10 days. Differentiation potential and gene expres- sion of 3, 6 and 12 months cryopreserved Lb-MSCs were unaltered. The results show that cryopreservation did not have an effect on the differentiation potential of human Lb-MSCs. Therefore, our work offers Lb- MSCs as clinically cells for treating osteo-diseases. Keywords Long bone Á Mesenchymal stem cells Á Cryopreservation Á Differentiation Á Cell banking Á Stem cell therapy S. Toosi Á F. Kalalinia Á J. Behravan (&) Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran e-mail: Behravanj@mums.ac.ir H. Naderi-Meshkin Á A. R. Bahrami Á A. Heirani-Tabasi Á M. Mirahmadi Stem Cell and Regenerative Medicine Research Group, Academic Center for Education, Culture and Research (ACECR), Khorasan Razavi Branch, Mashhad, Iran F. Kalalinia Medical Genetic Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran M. T. Pievandi Department of Orthopedic Surgery, Orthopedic and Trauma Research Center, Mashhad University of Medical Sciences, Mashhad, Iran H. Hosseinkhani School of Biomedical Engineering, National Yang Ming University, Taipei, Taiwan J. Behravan Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad, Iran 123 Cell Tissue Bank DOI 10.1007/s10561-017-9652-3