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Environmental regulation of notochordal gene expression in nucleus pulposus cells
Author(s) -
Rastogi Anshu,
Thakore Pratiksha,
Leung Aileen,
Benavides Magda,
Machado Monica,
Morschauser Michael A.,
Hsieh Adam H.
Publication year - 2009
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.21816
Subject(s) - notochord , microbiology and biotechnology , gene expression , biology , extracellular matrix , phenotype , chondrocyte , embryonic stem cell , population , cell , gene , chemistry , cartilage , anatomy , embryo , genetics , embryogenesis , demography , sociology
Cells of the nucleus pulposus (NP) in the intervertebral disc are derived directly from the embryonic notochord. In humans, a shift in NP cell population coincides with the beginning of age‐related changes in the extracellular matrix that can lead to spinal disorders. To begin identifying the bases of these changes, the manner by which relevant environmental factors impact cell function must be understood. This study investigated the roles of biochemical, nutritional, and physical factors in regulating immature NP cells. Specifically, we examined cell morphology, attachment, proliferation, and expression of genes associated with the notochord and immature NP (Sox9, CD24, and type IIA procollagen). Primary cells isolated from rat caudal discs were exposed to different media formulations and physical culture configurations either in 21% (ambient) or 2% (hypoxic) O 2 . As expected, cells in alginate beads retained a vacuolated morphology similar to chordocytes, with little change in gene expression. Interestingly, NP tissues not enzymatically digested were more profoundly influenced by oxygen. In monolayer, α‐MEM preserved vacuolated morphology, produced the highest efficiency of attachment, and best maintained gene expression. DMEM and Opti‐MEM cultures resulted in high levels of proliferation, but these appeared to involve small non‐vacuolated cells. Gene expression patterns for cells in DMEM monolayer cultures were consistent with chondrocyte de‐differentiation, with the response being delayed by hypoxia. Overall, results indicate that certain environmental conditions induce cellular changes that compromise the notochordal phenotype in immature NP. These results form the foundation on which the mechanisms of such changes can be elucidated. J. Cell. Physiol. 220: 698–705, 2009. © 2009 Wiley‐Liss, Inc.