caa a22 4500 44529227X CHVBK 20180317142532.0 cr unu---uuuuu 170323e20100101xx s 000 0 eng 10.1007/s11626-009-9238-x doi (NATIONALLICENCE)springer-10.1007/s11626-009-9238-x Osteogenesis in vitro: from pre-osteoblasts to osteocytes [Elektronische Daten] A contribution from the Osteobiology Research Group, The Pennsylvania State University [Venkatesh Krishnan, Ravi Dhurjati, Erwin Vogler, Andrea Mastro] Murine calvariae pre-osteoblasts (MC3T3-E1), grown in a novel bioreactor, proliferate into a mineralizing 3D osteoblastic tissue that undergoes progressive phenotypic maturation into osteocyte-like cells. Initially, the cells are closely packed (high cell/matrix ratio), but transform into a more mature phenotype (low cell/matrix ratio) after about 5mo, a process that recapitulates stages of bone development observed in vivo. The cell morphology concomitantly evolves from spindle-shaped pre-osteoblasts through cobblestone-shaped osteoblasts to stellate-shaped osteocyte-like cells interconnected by many intercellular processes. Gene-expression profiles parallel cell morphological changes, up-to-and-including increased expression of osteocyte-associated genes such as E11, DMP1, and sclerostin. X-ray scattering and infrared spectroscopy of contiguous, square centimeter-scale macroscopic mineral deposits are consistent with bone hydroxyapatite, showing that bioreactor conditions can lead to ossification reminiscent of bone formation. Thus, extended-term osteoblast culture (≤10mo) in a bioreactor based on the concept of simultaneous growth and dialysis captures the full continuum of bone development otherwise inaccessible with conventional cell culture, resulting in an in vitro model of osteogenesis and a source of terminally differentiated osteocytes that does not require demineralization of fully formed bone. The Society for In Vitro Biology, 2009 Osteocyte nationallicence Osteoblast nationallicence Bone formation nationallicence Bioreactor nationallicence Three-dimensional cell culture model nationallicence Krishnan Venkatesh Biochemistry and Molecular Biology, Pennsylvania State University, 431, South Frear, 16802, University Park, PA, USA aut Dhurjati Ravi Departments of Materials Science and Engineering, Pennsylvania State University, 16802, University Park, PA, USA aut Vogler Erwin Departments of Materials Science and Engineering, Pennsylvania State University, 16802, University Park, PA, USA aut Mastro Andrea Biochemistry and Molecular Biology, Pennsylvania State University, 431, South Frear, 16802, University Park, PA, USA aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/1(2010-01-01), 28-35 1071-2690 46:1<28 2010 46 11626 https://doi.org/10.1007/s11626-009-9238-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-009-9238-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Krishnan Venkatesh Biochemistry and Molecular Biology, Pennsylvania State University, 431, South Frear, 16802, University Park, PA, USA aut NATIONALLICENCE 700 1- Dhurjati Ravi Departments of Materials Science and Engineering, Pennsylvania State University, 16802, University Park, PA, USA aut NATIONALLICENCE 700 1- Vogler Erwin Departments of Materials Science and Engineering, Pennsylvania State University, 16802, University Park, PA, USA aut NATIONALLICENCE 700 1- Mastro Andrea Biochemistry and Molecular Biology, Pennsylvania State University, 431, South Frear, 16802, University Park, PA, USA aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/1(2010-01-01), 28-35 1071-2690 46:1<28 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer