In vitro transduktion humaner osteoblastarer zellpopulationen mit retroviralen vektoren

Translated title of the contribution: In vitro transduction of human osteoblastic cells using retroviral vectors

A. W A Baltzer, J. D. Whalen, T. Muzzonegro, H. I. Georgescu, P. D. Robbins, Christopher H Evans

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Objectives: The involvement of cytokines in degeneration and inflammation of human tissue is well established. Interleukin- 1 (IL-1) is a major agent in the pathophysiology of periarticular bone resorption in rheumatoid arthritis and in osteoporosis. Because the use of recombinant cytokines and growth factors is limited due to their short half lives, techniques are needed to get a permanent release of these therapeutic proteins. The rational of this study was to show that retroviral transduction of human osteoblastic cells is possible in vitro using the marker gene LacZ and the potentially therapeutic gene encoding for human interleukin-1 receptor antagonist protein (IL-1Ra). Different transduction techniques were combined to improve the rate of transduction in vitro. Methods: Osteoblastic cells were isolated from human spongious bone and cultured in vitro. The β- galactosidase (LacZ) gene and the cDNA of IL-1Ra were introduced into the isolated cells by retrovirus mediated gene transfer. LacZ activity was determined by Xga staining, IL-1Ra was measured quantitatively by ELISA. Results: The transfer of retroviral IL-1Ra led to IL-1Ra expression of 8614 to 10089 pg IRAP/50 000 cells/48 h. By combining different techniques to improve transduction, the X-gal staining established a rate of transduction of 60%. Conclusion: Our results demonstrate that retroviral transduction of human osteoblastic cells is possible in vitro, and leads to high levels of the synthesized transgene product. The rate of retroviral transduction can be accelerated in vitro.

Original languageGerman
Pages (from-to)88-94
Number of pages7
JournalZeitschrift fur Rheumatologie
Volume58
Issue number2
DOIs
StatePublished - Apr 1999
Externally publishedYes

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Interleukin 1 Receptor Antagonist Protein
Lac Operon
Galactosidases
Staining and Labeling
Cytokines
Retroviridae
Bone Resorption
Transgenes
Interleukin-1
Genes
Osteoporosis
Rheumatoid Arthritis
Intercellular Signaling Peptides and Proteins
Complementary DNA
Enzyme-Linked Immunosorbent Assay
In Vitro Techniques
Inflammation
Bone and Bones
Therapeutics
Proteins

Keywords

  • Gene therapy
  • Interleukin-1 receptor antagonist
  • Osteoblastic cells
  • Retrovirus

ASJC Scopus subject areas

  • Rheumatology

Cite this

In vitro transduktion humaner osteoblastarer zellpopulationen mit retroviralen vektoren. / Baltzer, A. W A; Whalen, J. D.; Muzzonegro, T.; Georgescu, H. I.; Robbins, P. D.; Evans, Christopher H.

In: Zeitschrift fur Rheumatologie, Vol. 58, No. 2, 04.1999, p. 88-94.

Research output: Contribution to journalArticle

Baltzer, A. W A ; Whalen, J. D. ; Muzzonegro, T. ; Georgescu, H. I. ; Robbins, P. D. ; Evans, Christopher H. / In vitro transduktion humaner osteoblastarer zellpopulationen mit retroviralen vektoren. In: Zeitschrift fur Rheumatologie. 1999 ; Vol. 58, No. 2. pp. 88-94.
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