title: Osteogenesis in vitro: from pre-osteoblasts to osteocytes: a contribution from the Osteobiology Research Group, The Pennsylvania State University.
abstract: 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 5 mo, 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 (< or =10 mo) 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.
authors.affiliation: Biochemistry and Molecular Biology, Pennsylvania State University, 431, South Frear, University Park, PA 16802, USA.
authors.forename: Venkatesh
authors.initials: V
authors.lastname: Krishnan
authors.forename: Ravi
authors.initials: R
authors.lastname: Dhurjati
authors.forename: Erwin A
authors.initials: EA
authors.lastname: Vogler
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Mastro
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D019149:Bioreactors; D002113:Calcification, Physiologic / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D048430:Cell Shape; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D051379:Mice; D018613:Microscopy, Confocal; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D010011:Osteocytes / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D010012:Osteogenesis / Q000502:physiology*; D010414:Pennsylvania; D010641:Phenotype; D012106:Research; D013997:Time Factors; D014495:Universities
chemical_list: D015415:Biomarkers