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Genetic marking with the ΔLNGFR‐gene for tracing goat cells in bone tissue engineering
Author(s) -
Kruyt M. C.,
Stijns M. M. C.,
Fedorovich N. E.,
de Bruijn J. D.,
van Blitterswijk C. A.,
Verbout A. J.,
Rozemuller H.,
Hagenbeek A.,
Dhert W. J. A.,
Martens A. C. M.
Publication year - 2004
Publication title -
journal of orthopaedic research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.041
H-Index - 155
eISSN - 1554-527X
pISSN - 0736-0266
DOI - 10.1016/j.orthres.2003.10.021
Subject(s) - stromal cell , bone marrow , tissue engineering , biology , microbiology and biotechnology , viral vector , in vivo , gene expression , immunology , gene , cancer research , recombinant dna , genetics
Abstract The use of bone marrow derived stromal cells (BMSC's) for bone tissue engineering has gained much attention as an alternative for autologous bone grafting. Little is known however, about the survival and differentiation of the cells, especially in the clinical application. The aim of this study was to develop a method to trace goat BMSC's in vivo. We investigated retroviral genetic marking, which allows stable expression of the label with cell division. Goat BMSC's were subjected to an amphotropic envelope containing a MoMuLV‐based vector expressing the human low affinity nerve growth factor receptor (ΔLNGFR). Labeling efficiency and effect on the cells were analyzed. Furthermore, transduced cells were seeded onto porous ceramic scaffolds, implanted subcutaneously in nude mice and examined after successive implantation periods. Flow cytometry indicated a transduction efficiency of 40–60%. Immunohistochemistry showed survival and subsequent bone formation of the gene‐marked cells in vivo. Besides, marked cells were also found in cartilage and fibrous tissue. These findings indicate the maintenance of the precursor phenotype following gene transfer as well as the ability of the gene to be expressed following differentiation. We conclude that retroviral gene marking with ΔLNGFR is applicable to trace goat BMSC's in bone tissue engineering research. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.