Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro

J. E. Onyia, B. Miller, J. Hulman, J. Liang, R. Galvin, C. Frolik, S. Chandrasekhar, A. K. Harvey, Joseph Bidwell, J. Herring, J. M. Hock

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

We have shown that intermittent parathyroid hormone (PTH) treatment targets proliferating cells in the primary spongiosa of trabecular bone of young rats, resulting in an increased number of osteoblasts. To further characterize these proliferating osteoprogenitor cells, bromodeoxyuridine (BrdUrd) incorporated in vivo, was used as a marker to identify and isolate cells for in vitro studies. Proliferating cells were labeled in vivo in young rats with BrdUrd and 24 h later were isolated by trypsinization of sections of the primary spongiosa of the distal femur metaphysis. Within 12 h of isolation, BrdUrd+ cells formed distinct foci containing 20-500 cells with fibroblast morphology. Stimulation of proliferation as determined by [3H]-thymidine incorporation was observed for these cells in response to fetal bovine serum, platelet derived growth factor, and transforming growth factor β-1 Neither insulin-like growth factor-1 (IGF-1) nor insulin stimulated proliferation PTH (1-34) and dexamethasone inhibited proliferation. The effects of PTH and dexamethasone were additive. Cells expressed the osteoblast phenotype as evidenced by synthesis of type I collagen, expression of high alkaline phosphatase activity, and production of increased intracellular cAMP in response to PTH (1-34). Confluent cell aggregates spontaneously formed mineralized nodules within 4-7 days, in the absence of inducers. These observations suggest that the primary spongiosa cells recapitulates the differentiation process in vitro in an accelerated fashion and may serve as a useful model to study osteoblast differentiation.

Original languageEnglish
Pages (from-to)93-100
Number of pages8
JournalBone
Volume20
Issue number2
DOIs
StatePublished - Feb 1997

Fingerprint

Phenotype
Parathyroid Hormone
Bromodeoxyuridine
Osteoblasts
Dexamethasone
In Vitro Techniques
Cell Separation
Platelet-Derived Growth Factor
Transforming Growth Factors
Somatomedins
Collagen Type I
Femur
Thymidine
Alkaline Phosphatase
Cell Differentiation
Fibroblasts
Insulin
Serum

Keywords

  • Osteoblast
  • Primary spongiosa
  • Proliferating cells
  • Rat

ASJC Scopus subject areas

  • Physiology
  • Hematology

Cite this

Onyia, J. E., Miller, B., Hulman, J., Liang, J., Galvin, R., Frolik, C., ... Hock, J. M. (1997). Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro. Bone, 20(2), 93-100. https://doi.org/10.1016/S8756-3282(96)00350-X

Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro. / Onyia, J. E.; Miller, B.; Hulman, J.; Liang, J.; Galvin, R.; Frolik, C.; Chandrasekhar, S.; Harvey, A. K.; Bidwell, Joseph; Herring, J.; Hock, J. M.

In: Bone, Vol. 20, No. 2, 02.1997, p. 93-100.

Research output: Contribution to journalArticle

Onyia, JE, Miller, B, Hulman, J, Liang, J, Galvin, R, Frolik, C, Chandrasekhar, S, Harvey, AK, Bidwell, J, Herring, J & Hock, JM 1997, 'Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro', Bone, vol. 20, no. 2, pp. 93-100. https://doi.org/10.1016/S8756-3282(96)00350-X
Onyia JE, Miller B, Hulman J, Liang J, Galvin R, Frolik C et al. Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro. Bone. 1997 Feb;20(2):93-100. https://doi.org/10.1016/S8756-3282(96)00350-X
Onyia, J. E. ; Miller, B. ; Hulman, J. ; Liang, J. ; Galvin, R. ; Frolik, C. ; Chandrasekhar, S. ; Harvey, A. K. ; Bidwell, Joseph ; Herring, J. ; Hock, J. M. / Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro. In: Bone. 1997 ; Vol. 20, No. 2. pp. 93-100.
@article{7b8f4f46057c48f18bf4d42d2d05cf95,
title = "Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro",
abstract = "We have shown that intermittent parathyroid hormone (PTH) treatment targets proliferating cells in the primary spongiosa of trabecular bone of young rats, resulting in an increased number of osteoblasts. To further characterize these proliferating osteoprogenitor cells, bromodeoxyuridine (BrdUrd) incorporated in vivo, was used as a marker to identify and isolate cells for in vitro studies. Proliferating cells were labeled in vivo in young rats with BrdUrd and 24 h later were isolated by trypsinization of sections of the primary spongiosa of the distal femur metaphysis. Within 12 h of isolation, BrdUrd+ cells formed distinct foci containing 20-500 cells with fibroblast morphology. Stimulation of proliferation as determined by [3H]-thymidine incorporation was observed for these cells in response to fetal bovine serum, platelet derived growth factor, and transforming growth factor β-1 Neither insulin-like growth factor-1 (IGF-1) nor insulin stimulated proliferation PTH (1-34) and dexamethasone inhibited proliferation. The effects of PTH and dexamethasone were additive. Cells expressed the osteoblast phenotype as evidenced by synthesis of type I collagen, expression of high alkaline phosphatase activity, and production of increased intracellular cAMP in response to PTH (1-34). Confluent cell aggregates spontaneously formed mineralized nodules within 4-7 days, in the absence of inducers. These observations suggest that the primary spongiosa cells recapitulates the differentiation process in vitro in an accelerated fashion and may serve as a useful model to study osteoblast differentiation.",
keywords = "Osteoblast, Primary spongiosa, Proliferating cells, Rat",
author = "Onyia, {J. E.} and B. Miller and J. Hulman and J. Liang and R. Galvin and C. Frolik and S. Chandrasekhar and Harvey, {A. K.} and Joseph Bidwell and J. Herring and Hock, {J. M.}",
year = "1997",
month = "2",
doi = "10.1016/S8756-3282(96)00350-X",
language = "English",
volume = "20",
pages = "93--100",
journal = "Bone",
issn = "8756-3282",
publisher = "Elsevier Inc.",
number = "2",

}

TY - JOUR

T1 - Proliferating cells in the primary spongiosa express osteoblastic phenotype in vitro

AU - Onyia, J. E.

AU - Miller, B.

AU - Hulman, J.

AU - Liang, J.

AU - Galvin, R.

AU - Frolik, C.

AU - Chandrasekhar, S.

AU - Harvey, A. K.

AU - Bidwell, Joseph

AU - Herring, J.

AU - Hock, J. M.

PY - 1997/2

Y1 - 1997/2

N2 - We have shown that intermittent parathyroid hormone (PTH) treatment targets proliferating cells in the primary spongiosa of trabecular bone of young rats, resulting in an increased number of osteoblasts. To further characterize these proliferating osteoprogenitor cells, bromodeoxyuridine (BrdUrd) incorporated in vivo, was used as a marker to identify and isolate cells for in vitro studies. Proliferating cells were labeled in vivo in young rats with BrdUrd and 24 h later were isolated by trypsinization of sections of the primary spongiosa of the distal femur metaphysis. Within 12 h of isolation, BrdUrd+ cells formed distinct foci containing 20-500 cells with fibroblast morphology. Stimulation of proliferation as determined by [3H]-thymidine incorporation was observed for these cells in response to fetal bovine serum, platelet derived growth factor, and transforming growth factor β-1 Neither insulin-like growth factor-1 (IGF-1) nor insulin stimulated proliferation PTH (1-34) and dexamethasone inhibited proliferation. The effects of PTH and dexamethasone were additive. Cells expressed the osteoblast phenotype as evidenced by synthesis of type I collagen, expression of high alkaline phosphatase activity, and production of increased intracellular cAMP in response to PTH (1-34). Confluent cell aggregates spontaneously formed mineralized nodules within 4-7 days, in the absence of inducers. These observations suggest that the primary spongiosa cells recapitulates the differentiation process in vitro in an accelerated fashion and may serve as a useful model to study osteoblast differentiation.

AB - We have shown that intermittent parathyroid hormone (PTH) treatment targets proliferating cells in the primary spongiosa of trabecular bone of young rats, resulting in an increased number of osteoblasts. To further characterize these proliferating osteoprogenitor cells, bromodeoxyuridine (BrdUrd) incorporated in vivo, was used as a marker to identify and isolate cells for in vitro studies. Proliferating cells were labeled in vivo in young rats with BrdUrd and 24 h later were isolated by trypsinization of sections of the primary spongiosa of the distal femur metaphysis. Within 12 h of isolation, BrdUrd+ cells formed distinct foci containing 20-500 cells with fibroblast morphology. Stimulation of proliferation as determined by [3H]-thymidine incorporation was observed for these cells in response to fetal bovine serum, platelet derived growth factor, and transforming growth factor β-1 Neither insulin-like growth factor-1 (IGF-1) nor insulin stimulated proliferation PTH (1-34) and dexamethasone inhibited proliferation. The effects of PTH and dexamethasone were additive. Cells expressed the osteoblast phenotype as evidenced by synthesis of type I collagen, expression of high alkaline phosphatase activity, and production of increased intracellular cAMP in response to PTH (1-34). Confluent cell aggregates spontaneously formed mineralized nodules within 4-7 days, in the absence of inducers. These observations suggest that the primary spongiosa cells recapitulates the differentiation process in vitro in an accelerated fashion and may serve as a useful model to study osteoblast differentiation.

KW - Osteoblast

KW - Primary spongiosa

KW - Proliferating cells

KW - Rat

UR - http://www.scopus.com/inward/record.url?scp=18844482573&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=18844482573&partnerID=8YFLogxK

U2 - 10.1016/S8756-3282(96)00350-X

DO - 10.1016/S8756-3282(96)00350-X

M3 - Article

C2 - 9028532

AN - SCOPUS:18844482573

VL - 20

SP - 93

EP - 100

JO - Bone

JF - Bone

SN - 8756-3282

IS - 2

ER -