Abstract
Macrophage-colony stimulating factor (M-CSF) has been reported to be required for the proliferation and differentiation of macrophages from hematopoietic progenitor cells. Recently, recombinant M-CSF (rM-CSF) became widely used as a biological research reagent in bone marrow stimulations, vaccine development, gene therapy approaches, and stem cell mobilization. rM-CSF is a glycoprotein that activates and enhances the differentiation and survival of macrophages, which play a key role in the osteoclastogenetic response. Here, we describe the construction of the gene encoding rM-CSF, its cloning, and expression in Escherichia coli, as well as the purification of rM-CSF protein, and its activity in a biological assay in mouse bone marrow cells. Our results show that the combination of experimental strategies employed to obtain recombinant rM-CSF can yield a biologically active protein, and may be useful when scaling-up production of other biologically similar proteins.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Rosen, C. J. (2013) American Society for Bone and Mineral Research: Primer on the metabolic bone diseases and disorders of mineral metabolism. 8th Ed., Wiley-Blackwell, Ames, Iowa.
Marino, S., J. G. Logan, D. Mellis, and M. Capulli (2014) Generation and culture of osteoclasts. BoneKEy Rep. 3: 570.
Suda, T., N. Udagawa, I. Nakamura, C. Miyaura, and N. Takahashi (1995) Modulation of osteoclast differentiation by local factors. Bone. 17: 87S–91S.
Tanaka, S., N. Takahashi, N. Udagawa, T. Tamura, T. Akatsu, E. R. Stanley, T. Kurokawa, and T. Suda (1993) Macrophage colony-stimulating factor is indispensable for both proliferation and differentiation of osteoclast progenitors. The J. Clin. Investigat. 91: 257–263.
Yoshida, H., S. Hayashi, T. Kunisada, M. Ogawa, S. Nishikawa, H. Okamura, T. Sudo, and L. D. Shultz (1990) The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene. Nature 345: 442–444.
Felix, R., M. G. Cecchini, and H. Fleisch (1990) Macrophage colony stimulating factor restores in vivo bone resorption in the op/op osteopetrotic mouse. Endocrinol. 127: 2592–2594.
Robb, L. (2007) Cytokine receptors and hematopoietic differentiation. Oncogene. 26: 6715–6723.
Sapi, E. and B. M. Kacinski (1999) The role of CSF-1 in normal and neoplastic breast physiology. 220: 1-8. Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine. New York, USA
Rettenmier, C. W., M. F. Roussel, and C. J. Sherr (1988) The colony-stimulating factor 1 (CSF-1) receptor (c-fms proto-oncogene product) and its ligand. J Cell Sci. 9: 27–44.
Kawasaki, E. S., M. B. Ladner, A. M. Wang, J. Van Arsdell, M. K. Warren, M. Y. Coyne, V. L. Schweickart, M. T. Lee, K. J. Wilson, and A. Boosman (1985) Molecular cloning of a complementary DNA encoding human macrophage-specific colonystimulating factor (CSF-1). Sci. 230: 291–296.
Ladner, M. B., G. A. Martin, J. A. Noble, D. M. Nikoloff, R. Tal, E. S. Kawasaki, and T. J. White (1987) Human CSF-1: Gene structure and alternative splicing of mRNA precursors. The EMBO J. 6: 2693–2698.
Wong, G. G., P. A. Temple, A. C. Leary, J. S. Witek-Giannotti, Y. C. Yang, A. B. Ciarletta, M. Chung, P. Murtha, R. Kriz, and R. J. Kaufman (1987) Human CSF-1: Molecular cloning and expression of 4-kb cDNA encoding the human urinary protein. Sci. 235: 1504–1508.
Pampfer, S., S. Tabibzadeh, F. C. Chuan, and J. W. Pollard (1991) Expression of colony-stimulating factor-1 (CSF-1) messenger RNA in human endometrial glands during the menstrual cycle: molecular cloning of a novel transcript that predicts a cell surface form of CSF-1. Mol. Endocrinol. 5: 1931–1938.
Nandi, S., M. P. Akhter, M. F. Seifert, X. M. Dai, and E. R. Stanley (2006) Developmental and functional significance of the CSF-1 proteoglycan chondroitin sulfate chain. Blood 107: 786–795.
Horiuchi, K., T. Miyamoto, H. Takaishi, A. Hakozaki, N. Kosaki, Y. Miyauchi, M. Furukawa, J. Takito, H. Kaneko, K. Matsuzaki, H. Morioka, C. P. Blobel, and Y. Toyama (2007) Cell surface colony-stimulating factor 1 can be cleaved by TNF-alpha converting enzyme or endocytosed in a clathrin-dependent manner. J. Immunol. 179: 6715–6724.
Yao, G. Q., J. J. Wu, B. H. Sun, N. Troiano, M. A. Mitnick, and K. Insogna (2003) The cell surface form of colony-stimulating factor-1 is biologically active in bone in vivo. Endocrinol. 144: 3677–3682.
Morohashi, T., V. A. Corboz, H. Fleisch, M. G. Cecchini, and R. Felix (1994) Macrophage colony-stimulating factor restores bone resorption in op/op bone in vitro in conjunction with parathyroid hormone or 1,25-dihydroxyvitamin D3. J. Bone and Mineral Res. 9: 401–407.
Koths, K. (1997) Structure-function studies on human macrophage colony-stimulating factor (M-CSF). Mol. Reprod. Dev. 46: 31–37.
Bessette, P. H., F. Aslund, J. Beckwith, and G. Georgiou (1999) Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm. Proc. Natl. Acad. Sci. USA. 96: 13703–13708.
Sohn, H., Y. Ko, M. Park, D. Kim, Y. L. Moon, Y. J. Jeong, H. Lee, Y. Moon, B. C. Jeong, O. Kim, and W. Lim (2015) Effects of light-emitting diode irradiation on RANKL-induced osteoclastogenesis. Lasers Surg. Med. 47: 745–755.
Zhang, P., J. Liu, Q. Xu, G. Harber, X. Feng, S. M. Michalek, and J. Katz (2011) TLR2-dependent modulation of osteoclastogenesis by Porphyromonas gingivalis through differential induction of NFATc1 and NF-kappaB. J. Biol. Chem. 286: 24159–24169.
Author information
Authors and Affiliations
Corresponding author
Additional information
HongMoon Sohn and Youngjong Ko contributed equally to this work.
Rights and permissions
About this article
Cite this article
Sohn, H., Ko, Y., Park, M. et al. Cloning and expression of recombinant macrophage-colony stimulating factor - A progressive strategy for economical production. Biotechnol Bioproc E 21, 446–452 (2016). https://doi.org/10.1007/s12257-016-0156-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-016-0156-y