Visualizing levels of osteoblast differentiation by a two-color promoter-GFP strategy: Type I collagen-GFPcyan and osteocalcin-GFPtpz

Bilic-Curcic, I, Kronenberg, M, Jiang, X, Bellizzi, J, Mina, M, Marijanovic, I, Gardiner, EM and Rowe, DW (2005) Visualizing levels of osteoblast differentiation by a two-color promoter-GFP strategy: Type I collagen-GFPcyan and osteocalcin-GFPtpz. Genesis, 43 2: 87-98. doi:10.1002/gene.20156


Author Bilic-Curcic, I
Kronenberg, M
Jiang, X
Bellizzi, J
Mina, M
Marijanovic, I
Gardiner, EM
Rowe, DW
Title Visualizing levels of osteoblast differentiation by a two-color promoter-GFP strategy: Type I collagen-GFPcyan and osteocalcin-GFPtpz
Journal name Genesis   Check publisher's open access policy
ISSN 1526-954X
Publication date 2005-01-01
Sub-type Article (original research)
DOI 10.1002/gene.20156
Volume 43
Issue 2
Start page 87
End page 98
Total pages 12
Editor R. Behringer
T. Magnuson
Place of publication USA
Publisher John Wiley & Sons, Inc.
Language eng
Subject C1
320204 Immunogenetics
730305 Diagnostic methods
Abstract A 3.9 kb DNA fragment of human osteocalcin promoter and 3.6 kb DNA fragment of the rat collagen type1a1 promoter linked with visually distinguishable GFP isomers, topaz and cyan, were used for multiplex analysis of osteoblast lineage progression. Three patterns of dual transgene, expression can be appreciated in primary bone cell cultures derived from the transgenic mice and by histology of their corresponding bones. Our data support the interpretation that strong pOBCol3.6GFPcyan alone is found in newly formed osteoblasts, while strong pOBCol3.6GFPcyan and hOC-GFPtpz are present in osteoblasts actively making a new matrix. Osteoblasts expressing strong hOC-GFPtpz and weak pOBCol3.6GF-Pcyan are also present and may or may not be producing mineralized matrix. This multiplex approach reveals the heterogeneity within the mature osteoblast population that cannot be appreciated by current histological methods. It should be useful to identify and isolate populations of cells within an osteoblast lineage as they progress through stages of differentiation.
Keyword Developmental Biology
Genetics & Heredity
Gfp
Osteocalcin Promoter
Osteoblast Differentiation
Type I Collagen Promoter
Primary Osteoblast Culture
Bone Histology
Gene-expression
Transgenic Mice
Murine Bone
Insitu Hybridization
Alkaline-phosphatase
Monoclonal-antibody
Progenitor Cells
Messenger-rna
Lineage
Cultures
Q-Index Code C1
Institutional Status Non-UQ

 
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Created: Wed, 15 Aug 2007, 16:49:36 EST