Metabolic and histological analysis of mesenchymal stem cells grown in 3-D hyaluronan-based scaffolds
Sheep mesenchymal stem cells (MSCs) were isolated and expanded using the principle of plastic adherence. Their identity as progenitor cells was confirmed by induction along the osteoblastic lineage using osteogenic supplements and observation of calcific deposits by von Kossa staining. MSCs were seeded onto two types of hyaluronan-based cylindrical scaffolds in high concentrations and cultured for varying time points up to three weeks. Culture medium was supplied using the following conditions: statically, on a shaker, by stirring with a magnetic stirrer or by perfusion in a tubular flow circuit. Total cell metabolism was assessed by MTT assay and the quality of cell coverage and matrix formation observed by SEM and histological analysis of thin sections of the constructs. Perfusion culture was established as the most appropriate culturing conditions, with cell metabolism increasing by approximately 300% over three weeks. The coverage of the scaffold surface was very good and the deposition of collagenous matrix was superior in these conditions compared to the static and other dynamic culture conditions.
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- 1.R. Langer and J. Vacanti, Science 260 (1993) 920.Google Scholar
- 2.L. A. Solchaga, B. Johnstone, J. U. Yoo, V. M. Goldberg and A. I. Caplan, Cell Transplant. 8 (1999) 511.Google Scholar
- 3.B. Johnstone, T. M. Hering, A. I. Caplan, V. M. Goldberg and J. U. Yoo, Exp. Cell Res. 238 (1998) 265.Google Scholar
- 4.A. I. Caplan, Clin. Plas. Surg. 21 (1994) 429.Google Scholar
- 5.A. I. Caplan, J. Orthop. Res. 9 (1991) 641.Google Scholar
- 6.L. E. Freed and G. Vunjak-Novakovic, Adv. Drug Deliv. Rev. 33 (1998) 15.Google Scholar