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Abstract westfield veterinary group Full-Text PDF Full-Text HTML Full-Text westfield veterinary group ePUB Linked References How to Cite this Article Complete Special Issue Stem Cells International Volume 2014 (2014), Article ID 276862, 6 pages http://dx.doi.org/10.1155/2014/276862
Fatty bone marrow (BM) and defective hematopoiesis are a pathologic hallmark of aplastic anemia (AA). We have investigated adipogenic and osteogenic potential of BM mesenchymal stem cells (BM-MSC) in 10 AA patients (08 males and 02 females) with median age of 37 years (range: 06 to 79 years) and in the same number westfield veterinary group of age and sex matched westfield veterinary group controls. It was observed westfield veterinary group that BM-MSC westfield veterinary group of AA patients westfield veterinary group had a morphology, phenotype, and osteogenic differentiation potential similar to control subjects but adipocytes differentiated from AA BM-MSC had a higher density and larger size of lipid droplets and they expressed significantly higher levels of adiponectin and FABP4 genes and proteins as compared to control BM-MSC ( for both). Thus our data shows that AA BM-MSC have enhanced westfield veterinary group adipogenicity, which may have an important implication in the pathogenesis of the disease. 1. Introduction
Aplastic anemia (AA) is a bone marrow westfield veterinary group (BM) failure syndrome characterized by a fatty BM and peripheral westfield veterinary group pancytopenia. Defects in hematopoietic westfield veterinary group stem cells (HSC) as well in the BM stroma have been implicated in the pathogenesis of AA but the exact cause of the disease is still obscure [ 1 ]. We have previously shown in AA an increased apoptosis of BM cells [ 2 ], IL-8 levels [ 3 ], and expression of interferon- λ and tumor necrosis westfield veterinary group factor- α in BM T-cells as well as their increased levels in BM plasma [ 4 ]. These studies point towards the role of immune mechanisms and bone marrow microenvironment in the pathogenesis of the disease. Mesenchymal stem cells (MSC) are the key stem cells of the BM microenvironment that give rise to different stromal cell types including adipocytes, westfield veterinary group osteoblasts, endothelial cells, and stromal fibroblasts and maintain hematopoietic homeostasis in the marrow by cell-cell contact and by producing various hematopoietic cytokines and growth factors [ 5 , 6 ]. The AA BM-MSC have been shown to have an abnormal gene expression profile [ 7 ] and abnormal immunological properties [ 8 , 9 ] indicating a BM-MSC dysfunction in AA. It is also recently reported that adipocytes present in the BM suppress HSC maturation and differentiation and an imbalance between adipogenic and osteogenic differentiation westfield veterinary group of MSC may substantially influence hematopoiesis [ 10 12 ]. Thus, in order to further explore the role of BM-MSC in the disease, we have evaluated their adipogenic and osteogenic differentiation potential in patients with AA. 2. Materials and Methods 2.1. Subjects and Culture and Phenotypic Characterization westfield veterinary group of BM-MSC
Ten AA patients [ 12 ], 08 males and 02 females with median age of 37 years (range: 6 to 79 years), and the same number of age and sex matched controls were recruited in the study. After informed consent, 5 mL of BM was aspirated from the posterior superior iliac crest of each subject and BM-MSC westfield veterinary group were isolated, cultured and phenotypically characterized as per the standard protocol established in the lab [ 13 ]. The cells of 3rd passage were used in the experiments. 2.2. Adipogenic Differentiation westfield veterinary group
BM-MSC in 3rd passage were treated with adipogenic medium consisting of DMEM medium (Invitrogen) containing 10% FBS (Hyclone), 500 mM IBMX, 1 mM dexamethasone, 10 mg/mL insulin, and 100 mM indomethacin (adipogenesis kit, Chemicon). After 18 days, the cells were fixed and stained with oil red O stain to visualize the fat droplets in the cells. 2.3. Osteogenic Differentiation
BM-MSC in 3rd passage were treated with osteogenic medium consisting of DMEM medium (Gibco-Invitrogen) containing 10% FBS (Hyclone), 1 mM dexamethasone, 10 mg/mL glyceraldehyde 3-phosphate, and 0.1 mM ascorbic acid (osteogenesis kit, Chemicon). After 21 days, the cells were fixed with 4% paraformaldehyde and stained with alizarin red stain to visualize mineralization. 2.4. Reverse-Transcription Polymerase Chain Reaction (RT-PCR) westfield veterinary group
Expression of adiponectin, fatty acid binding protein 4 (FABP4) and osteopontin was done by RT-PCR. Total RNA of BM-MSC of AA patients and controls westfield veterinary group was extracted using RNeasy Mini RNA isolation kit (Invitrogen). One μ g of total RNA was reverse transcribed into cDNA using random hexamers (Invitrogen). The gene primers (MWG Biotech, http://www.mwg-biotech.com/ ) used were