PCR detection of cytokines in normal human and pagetic osteoblast-like cells.
We investigated the expression of cytokine transcripts in osteoblast-like cells derived from explants of pagetic and normal bone. A reverse transcription-linked PCR was used that allowed the simultaneous analysis of a range of cytokines. Normal osteoblast-like cells were found to contain the transcripts for IL-1 beta, IL-6, and TGF-beta 1. For the first time we detected in bone cells the two other mammalian isoforms of TGF-beta, beta 2, and beta 3. Furthermore, we have also identified mRNA for IL-3 and the novel chemotactic factor, IL-8. Using this sensitive technique it was not possible to detect mRNA for IL-1 alpha, IL-2, IL-4, IL-5, IL-7, TNF-alpha, or interferon-gamma. The human osteosarcoma cell line Saos-2 also showed a similar pattern of expression of these cytokines to primary osteoblast-like cells, with the exception that TNF-alpha was also identified. Cells isolated from pagetic bone showed essentially the same profile of cytokine expression as normal bone except that TNF-alpha was also detected in two of four samples. The cytokine profile of successive populations of cells harvested from one explant culture at 9, 22, and 57 days showed a consistent pattern of cytokine expression, demonstrating the phenotypic stability of the osteoblast-like cells in long-term cultures.
Birch
M A
MA
Department of Human Anatomy and Cell Biology, University of Liverpool, England.
Ginty
A F
AF
Walsh
C A
CA
Fraser
W D
WD
Gallagher
J A
JA
Bilbe
G
G
D001483:Base Sequence; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D010001:Osteitis Deformans / Q000378:metabolism*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014158:Transcription, Genetic; D015291:Transforming Growth Factors / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
D016207:Cytokines; D018076:DNA, Complementary; D007378:Interleukins; D012333:RNA, Messenger; D015291:Transforming Growth Factors