CELLULAR & MOLECULAR BIOLOGY LETTERS http://www.cmbl.org.pl Received: 09 February 2015 Volume 20 (2015) pp 1-… Final form accepted: 12 June 2015 DOI: 10.1515/cmble-2015-0029 Published online: © 2015 by the University of Wrocław, Poland * Author for correspondence. Email: krzysztof.marycz@up.wroc.pl Abbreviations used: ALP – alkaline phosphatase; ASCs – multipotent stromal cells derived from adipose tissue; BM – bone marrow; BMPs – bone morphogenetic proteins; BMSCs – multipotent stromal cells derived from bone marrow; CerK – ceramide kinase; C1P – ceramide-1-phosphate; MSCs – mesenchymal stem cells; MVs – membrane-derived microvesicles; OCN – osteocalcin; OPN – osteopontin; OPs – osteoclast precursors; pNPP – p-nitrophenyl phosphate; SEM – scanning electron microscopy; SEM-EDX – scanning electron microscopy with energy dispersive X-ray analysis; SPHK – sphingosine kinase; S1P – sphingosine-1-phosphate; S1PRs – G-protein-coupled receptors Research article THE EFFECT OF THE BIOACTIVE SPHINGOLIPIDS S1P AND C1P ON MULTIPOTENT STROMAL CELLS – NEW OPPORTUNITIES IN REGENERATIVE MEDICINE KRZYSZTOF MARYCZ 1, 2, *, AGNIESZKA ŚMIESZEK 1, 2 , MARTA JELEŃ 2 , KLAUDIA CHRZĄSTEK 2 , JAKUB GRZESIAK 2 and JUSTYNA MEISSNER 2 1 Wrocław Research Centre EIT+, Stabłowicka 147, 54-066 Wrocław, Poland, 2 University of Environmental and Life Sciences Wroclaw, Faculty of Biology, Kożuchowska 5b, 50-631 Wrocław, Poland Abstract: Sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) belong to a family of bioactive sphingolipids that act as important extracellular signaling molecules and chemoattractants. This study investigated the influence of S1P and C1P on the morphology, proliferation activity and osteogenic properties of rat multipotent stromal cells derived from bone marrow (BMSCs) and subcutaneous adipose tissue (ASCs). We show that S1P and C1P can influence mesenchymal stem cells (MSCs), each in a different manner. S1P stimulation promoted the formation of cellular aggregates of BMSCs and ASCs, while C1P had an effect on the regular growth pattern and expanded intercellular connections, thereby increasing the proliferative activity. Although osteogenic differentiation of MSCs was enhanced by the addition of S1P, the effectiveness of osteoblast differentiation was more evident in BMSCs, particularly when biochemical and molecular marker levels were considered. The results of the functional osteogenic differentiation assay, which includes an evaluation of the efficiency of extracellular matrix mineralization (SEM-EDX), revealed the formation of numerous mineral aggregates in BMSC cultures stimulated with S1P. Our data demonstrated that in an appropriate combination, the bioactive Brought to you by | New York University Bobst Library Technical Services Authenticated Download Date | 6/30/15 7:48 AM