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Recombinant DNA technology has provided tools to introduce exogenous DNA into cultured mammalian cells. In cartilage tissue engineering, these methods can be used to study the effect of single genes on the regulation of chondrogenesis or to enhance structural and functional properties of engineered cartilage [Madry et al., 2002]. The techniques of transferring DNA molecules to cells can be divided into nonviral methods (including chemical, mechanical, and electrical techniques) and viral methods. The rst critical step is the delivery of a gene of interest to the chondrocyte. For the purposes of cartilage tissue engineering, this can be achieved by two different strategies. First, the DNA may be introduced into component chondrocytes before seeding them into a scaffold. The second approach would be to transfer the DNA at a later time point directly into tissue-engineered cartilaginous constructs. This step requires the transfection of chondrocytes within their native matrix in situ and can be performed ef ciently by using recombinant adeno-associated virus (rAAV) vectors [Madry et al., 2003b]. In general, an adequate delivery system requires (1) a high ef ciency of transmission, (2) a desired level and length of gene expression, and (3) biological safety during the course of the transgene expression. Recombinant genes have been ef ciently introduced into isolated articular chondrocytes by virus-based carriers [Madry et al., 2003b; Baragi et al., 1995; Doherty et al., 1998; Ikeda et al., 2000; Smith et al., 2000; Brower-Toland et al., 2001] and nonviral synthetic methods. Lipid-based vectors in particular offer many advantages [Madry et al., 2002; Kim et al., 1996; Madry and Trippel, 2000]. They are relatively easy to prepare and to handle, do not restrict the length of the DNA molecule of interest that is to be transferred, and very rarely elicit an immune response [Fogler et al., 1987]. The lipid-based gene transfer protocol described in this chapter has been optimized for overexpressing recombinant DNA in primary articular chondrocytes.
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n k S:|S|=k
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A message pops up to inform you that the defragmentation is complete. To read a detailed report about what occurred, click the View Report button. If you are not interested in seeing this report, simply click the Close button. Turn on your antivirus software by right-clicking its icon near the Windows clock and selecting Enable or Restore. If the antivirus icon is not there, then you must manually restart the program by doing the following: a. Click the Start button in the lower-left corner of Windows. b. Click All Programs. c. Select the folder containing the name of your antivirus software. For example, if you use Norton AntiVirus, then select the Norton AntiVirus folder. d. Click the shortcut to launch the program.
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Computation of Spearman correlation i,h 2,1 3,1 4,1 5,1 3,2 4,2 5,2 4,3 5,3 5,4 (qix qhx) 4 1 3 2 3 1 2 2 1 1 (qiy qhy) 1 2 3.5 3.5 1 2.5 2.5 1.5 1.5 0 Sums: rs = (qix qhx) (qiy qhy) 4 2 10.5 7 3 2.5 5 3 1.5 0 17.5 0.359 (qix qhx)2 16 1 9 4 9 1 4 4 1 1 50 (Equation 25.7) (qiy qhy)2 1 4 12.25 12.25 1 6.25 6.25 2.25 2.25 0 47.5
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