期刊
GENESIS
卷 43, 期 2, 页码 87-98出版社
WILEY
DOI: 10.1002/gene.20156
关键词
GFP; osteocalcin promoter; osteoblast differentiation; type I collagen promoter; primary osteoblast culture; bone histology
资金
- NIAMS NIH HHS [R01 AR43457] Funding Source: Medline
- NIDDK NIH HHS [U01 DK63478] Funding Source: Medline
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.
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