Osteogenesis in vitro: From pre-osteoblasts to osteocytes: A contribution from the osteobiology research group, the Pennsylvania state university

Venkatesh Krishnan, Ravi Dhurjati, Erwin A. Vogler, Andrea Marie Mastro

Research output: Contribution to journalArticle

20 Citations (Scopus)

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 (≤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.

Original languageEnglish (US)
Pages (from-to)28-35
Number of pages8
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume46
Issue number1
DOIs
StatePublished - Jan 1 2010

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Osteocytes
Osteoblasts
Osteogenesis
Research
Bioreactors
Bone Development
Bone and Bones
Durapatite
In Vitro Techniques
Transcriptome
Skull
Minerals
Dialysis
Spectrum Analysis
Cell Culture Techniques
X-Rays
Phenotype
Growth
Genes

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology

Cite this

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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 (≤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.",
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Osteogenesis in vitro : From pre-osteoblasts to osteocytes: A contribution from the osteobiology research group, the Pennsylvania state university. / Krishnan, Venkatesh; Dhurjati, Ravi; Vogler, Erwin A.; Mastro, Andrea Marie.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 46, No. 1, 01.01.2010, p. 28-35.

Research output: Contribution to journalArticle

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