Gene expression profiling of bone marrow mesenchymal stem cells from Osteogenesis Imperfecta patients during osteoblast differentiation Carla Martins Kaneto a, b, * , Patrícia S. Pereira Lima c , Karen Lima Prata d , Jane Lima dos Santos b , Jo ~ ao Monteiro de Pina Neto a , Rodrigo Alexandre Panepucci d , Houtan Noushmehr a , Dimas Tadeu Covas d , Francisco Jos  e Alburquerque de Paula e , Wilson Araújo Silva Jr. a, d a Department of Genetics, Ribeir~ ao Preto Medical School, Universidade de S~ ao Paulo, Ribeir~ ao Preto, S~ ao Paulo, Brazil b Department of Biological Science, Santa Cruz State University, Ilh eus, BA, Brazil c Department of Natural Science, Bahia State University, Vit oria da Conquista, Bahia, Brazil d Regional Blood Center of Ribeir~ ao Preto and National Institute of Science and Technology in Cell Therapy, Ribeir~ ao Preto, Brazil e Department of Clinical Medicine, Ribeir~ ao Preto Medical School, Universidade de S~ ao Paulo, Ribeir~ ao Preto, S~ ao Paulo, Brazil article info Article history: Received 14 September 2016 Received in revised form 22 March 2017 Accepted 5 April 2017 Available online xxx Keywords: Osteogenesis Imperfecta Bone marrow mesenchymal stem cell Osteogenic differentiation Gene expression abstract Mesenchymal stem cells (MSCs) are precursors present in adult bone marrow that are able to differ- entiate into osteoblasts, adipocytes and chondroblasts that have gained great importance as a source for cell therapy. Recently, a number of studies involving the analysis of gene expression of undifferentiated MSCs and of MSCs in the differentiation into multiple lineage processes were observed but there is no information concerning the gene expression of MSCs from Osteogenesis Imperfecta (OI) patients. Osteogenesis Imperfecta is characterized as a genetic disorder in which a generalized osteopenia leads to excessive bone fragility and severe bone deformities. The aim of this study was to analyze gene expression profile during osteogenic differentiation from BMMSCs (Bone Marrow Mesenchymal Stem Cells) obtained from patients with Osteogenesis Imperfecta and from control subjects. Bone marrow samples were collected from three normal subjects and five patients with OI. Mononuclear cells were isolated for obtaining mesenchymal cells that had been expanded until osteogenic differentiation was induced. RNA was harvested at seven time points during the osteogenic differentiation period (D0, Dþ1, Dþ2, Dþ7, Dþ12, Dþ17 and Dþ21). Gene expression analysis was performed by the microarray technique and identified several differentially expressed genes. Some important genes for osteoblast differentiation had lower expression in OI patients, suggesting a smaller commitment of these patient's MSCs with the osteogenic lineage. Other genes also had their differential expression confirmed by RT-qPCR. An increase in the expression of genes related to adipocytes was observed, suggesting an increase of adipogenic differentiation at the expense osteogenic differentiation. © 2017 Published by Elsevier Masson SAS. 1. Introduction Mesenchymal stromal cells or mesenchymal stem cells (MSCs) were first described by Fridensteins et al., in 1976, as clonal cells adherent to plastic and as a source of osteoblast, adipogenic and chondrogenic lineages. Bone marrow is the main source of MSCs but these cells can also be obtained from other tissues. There is a growing number of reports describing the obtaining of these cells from adipose tissue (Gronthos et al., 2001), umbilical cord blood (Igura et al., 2004), amniotic fluid (Tsai et al., 2004), peripheral blood (Zvaifler et al., 2000), fetal liver (Campagnoli et al., 2015), lung (Foundation, 2003) and even the pulp of deciduous teeth (Miura et al., 2003). MSCs are a heterogeneous population of cells in terms of morphology, physiology and expression of surface anti- gens. To date, a specific marker has not been identified for the * Corresponding author. Departamento de Ci^ encias Biol ogicas e DCB, Uni- versidade Estadual de Santa Cruz e UESC - Ilh eus, Rodovia Jorge Amado, Km16, 45662-900, Ilh eus, Bahia, Brazil. E-mail address: cmkaneto@uesc.br (C.M. Kaneto). Contents lists available at ScienceDirect European Journal of Medical Genetics journal homepage: http://www.elsevier.com/locate/ejmg http://dx.doi.org/10.1016/j.ejmg.2017.04.003 1769-7212/© 2017 Published by Elsevier Masson SAS. European Journal of Medical Genetics xxx (2017) 1e9 Please cite this article in press as: Kaneto, C.M., et al., Gene expression profiling of bone marrow mesenchymal stem cells from Osteogenesis Imperfecta patients during osteoblast differentiation, European Journal of Medical Genetics (2017), http://dx.doi.org/10.1016/ j.ejmg.2017.04.003