Microdialysis for monitoring the process of functional tissue culture.
Continuous monitoring is important during tissue culture. However, there are still technical difficulties in monitoring the internal status of cells or tissues. In this paper, microdialysis is adopted to monitor functional tissue growth in a bioreactor. Explanted bovine caudal intervertebral disc (IVD) was used as the test tissue. A microdialysis membrane probe of 100 kDa molecular weight cut-off was employed and in situ calibration methods with phenol red and fluorescent 40 kDa dextran were developed to measure the relative recovery of the solute of interest, and membrane fouling, respectively. Tissue metabolism was monitored successfully. At the same time soluble macromolecules were picked up by the probe and were detected and quantified by Fast Protein Liquid Chromatography (FPLC) and/or Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). These proteins were believed to be associated with biofunction of engineered tissue. Monitoring of phenol red content in the dialysate indicated that there was no significant fouling of the membrane probe during a 7-day culture period and the Relative Recovery of macromolecules of interests remained roughly 9%. We concluded that microdialysis could be used to sample a wide range of molecular species released during cell metabolism and extracellular matrix turnover, which were direct or indirect indications of cell and tissue functions. The application of the developed system could be extended to monitor tissue repair in vivo, and the development of the engineered tissue.
Li
Z
Z
Department of Engineering Science, University of Oxford, Oxford - UK.
Boubriak
O A
OA
Urban
J P
JP
Cui
Z F
ZF
D000818:Animals; D002417:Cattle; D017551:Microdialysis; D046509:Tissue Culture Techniques / Q000379:methods*