Volume 7, Issue 5, Pages (January 2005)

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Volume 7, Issue 5, Pages 447-455 (January 2005) Anterior cruciate ligament constructs fabricated from human mesenchymal stem cells in a collagen type I hydrogel U. Nöth, MD, K. Schupp, A. Heymer, S. Kall, F. Jakob, N. Schütze, B. Baumann, T. Barthel, J. Eulert, C. Hendrich Cytotherapy Volume 7, Issue 5, Pages 447-455 (January 2005) DOI: 10.1080/14653240500319093 Copyright © 2005 International Society for Cellular Therapy Terms and Conditions

Figure 1 Fabrication method of ACL constructs from a MSC–collagen gel suspension and bone cylinders. (a) Syringe filled with a bone cylinder on the bottom. (b) Filling of the syringe with the collagen gel containing MSC. (c) Final molded ACL construct with one bone cylinder on the bottom and one at the top of the syringe. Note that for later fixation in the bioreactor both cylinders were provided with a non-biodegradable wire. Cytotherapy 2005 7, 447-455DOI: (10.1080/14653240500319093) Copyright © 2005 International Society for Cellular Therapy Terms and Conditions

Figure 2 Bioreactor for cyclic stretching. (a) Schematic illustration of the bioreactor. (b) ACL construct attached to the mobile (top) and the fixed anchor with non-biodegradable wires. (c) Photograph of the bioreactor filled with medium and the ACL construct before starting the stretching experiment. Cytotherapy 2005 7, 447-455DOI: (10.1080/14653240500319093) Copyright © 2005 International Society for Cellular Therapy Terms and Conditions

Figure 3 Histochemical analysis of stretched ACL constructs (stretched) after 14 days compared with non-stretched controls (Ctrl) and a human ACL (human ACL). (a–c) H&E staining, (d–f) Masson-Goldner staining and (g–i) Azan staining. Bars = 100µm. Cytotherapy 2005 7, 447-455DOI: (10.1080/14653240500319093) Copyright © 2005 International Society for Cellular Therapy Terms and Conditions

Figure 4 Immunohistochemical analysis of stretched ACL constructs (stretched) after 14 days compared with non-stretched controls (Ctrl) and a human ACL (human ACL). (a–c) Staining for collagen type I (Coll I), (d–f) staining for collagen type III (Coll III), (g–i) staining for elastin and (j–l) staining for fibronectin (FN). Bars = 100µm. Cytotherapy 2005 7, 447-455DOI: (10.1080/14653240500319093) Copyright © 2005 International Society for Cellular Therapy Terms and Conditions

Figure 5 RT-PCR analysis of ligament-specific marker gene expression of constructs stretched for 14 days (St) compared with non-stretched controls (Ctrl). Coll I, collagen type I; Coll III, collagen type III; FN, fibronectin. Cytotherapy 2005 7, 447-455DOI: (10.1080/14653240500319093) Copyright © 2005 International Society for Cellular Therapy Terms and Conditions