Abstract
Laser capture microdissection (LCM) method allows selection of individual or clustered cells from intact tissues. This technology enables one to pick cells from tissues that are difficult to study individually, sort the anatomical complexity of these tissues, and make the cells available for molecular analyses. Following the cells' extraction, the nucleic acids and proteins can be isolated and used for multiple applications that provide an opportunity to uncover the molecular control of cellular fate in the natural microenvironment.
Utilization of LCM for the molecular analysis of cells from skeletal tissues will enable one to study differential patterns of gene expression in the native intact skeletal tissue with reliable interpretation of function for known genes as well as to discover novel genes. Variability between samples may be caused either by dif ferences in the tissue samples (different areas isolated from the same section) or some variances in sample handling. LCM is a multi-task technology that combines histology, microscopy work, and dedicated molecular biology. The LCM application will provide results that will pave the way toward high throughput profiling of tissue-specific gene expression using Gene Chip arrays. Detailed description of in vivo molecular pathways will make it possible to elaborate on control systems to apply for the repair of genetic or metabolic diseases of skeletal tissues.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
1. Friedenstein, A. J., Piatetzky, S., II, Petrakova, K. V. (1966) Osteogenesis in transplants of bone marrow cells. J Embryol Exp Morphol 16, 381–390.
2. Benayahu, D. (2000) The hematopoietic microenvironment: the osteogenic compartment of bone marrow: cell biology and clinical application. Hematology 4, 427–435.
3. Jiang, Y. , Jahagirdar, B. N., Reinhardt, R. L., et al. (2002) Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418, 41–49.
4. Pittenger, M. F., Mackay, A. M., Beck, S. C., et al. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147.
5. Benayahu D, Akavia UD, Shur I. (2007). Differentiation of bone marrow stroma-derived mesenchymal cells. Current Medical Chemistry. 14 (2):173–9.
6. Syed, F., Khosla, S. (2005) Mechanisms of sex steroid effects on bone. Biochem Biophys Res Commun 328, 688–696.
7. Benayahu, D., Shur, I., Ben-Eliyahu, S. (2000) Hormonal changes affect the bone and bone marrow cells in a rat model. J Cell Biochem 79, 407–415.
8. Compston, J. (2001) Sex steroids and bone. Physiol Rev 81, 419–447.
9. Locklin, R., Williamson, M., Beresford, J., et al. (1995) In vitro effects of growth factors and dexamethasone on rat marrow stromal cells. Clin Orthop 313, 27–35.
10. Benayahu, D., Akavia, U. D., Socher, R., Shur, I. (2005) Gene expression in skeletal tissues: application of laser capture microdissection. J Microsc 220(Pt 1), 1–8.
11. Benayahu D, Socher R, Shur I. 2007. Laser Capture Microdissection of Bone Cells. Methods in Molecular Biology/Molecular Medicine Osteoporosis Editor: Jennifer Westendorf published by Humana Press, USA.
12. Shur, I., Benayahu, D. (2005) Characterization and functional analysis of CReMM, a novel chromodomain helicase DNA-binding protein. J Mol Biol 352, 646–655.
13. Shur, I., Socher, R, Benayahu, D. (2006) In vivo association of CReMM/CHD9 with promot ers in osteogenic cells. J Cell Physiol 207, 374–378.
14. Shur, I., Salomon, R., Benayahu, D. (2006) Dynamic interactions of chromatin-related mes enchymal modulator, a chromodomain helicase-DNA-binding protein, with promoters in osteoprogenitors. Stem Cells 24, 1288–1293.
15. Marom, R., Shur, I., Hager, G. L., Benayahu, D. (2006) Expression and regulation of CReMM, a chromodomain helicase-DNA-binding (CHD), in marrow stroma derived osteo progenitors. J Cell Physiol 207(3), 628–35.
Acknowledgments
This study was supported by funding from Ramot at Tel Aviv University and by the Chief Scientist of Ministry of Commerce and consortium “Bereshit” and the CellProm program from the EEC 6th framework to D.B.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Benayahu, D., Socher, R., Shur, I. (2008). Application of the Laser Capture Microdissection Technique for Molecular Definition of Skeletal Cell Differentiation In Vivo . In: Westendorf, J.J. (eds) Osteoporosis. Methods In Molecular Biology™, vol 455. Humana Press. https://doi.org/10.1007/978-1-59745-104-8_15
Download citation
DOI: https://doi.org/10.1007/978-1-59745-104-8_15
Publisher Name: Humana Press
Print ISBN: 978-1-58829-828-7
Online ISBN: 978-1-59745-104-8
eBook Packages: Springer Protocols