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Optimization of Flexor Tendon Tissue Engineering: Human Adipoderived Stem Cell-Tenocyte Co-Cultures for Seeding of an Acellularized Tendon Scaffolds
Armin Kraus, MD1; Colin Woon, MD1; Shyam S. Raghavan, BS1; Hung Pham, BS1; Kai Megerle, MD1; Matthew Seung Suk Choi, MD2; James Chang, MD1
1Plastic & Reconstructive Surgery, Stanford University, Stanford, CA; 2Plastic & Reconstructive Surgery, Hanyang University Guri Hospital, Guri, South Korea

Introduction: Complex hand injuries often require multiple tendon grafting. The supply of autologous grafts is limited, so that there is a demand for artificial tendon grafts. Seeding acellularized tendons with cells is an approach to provide grafts with favorable biomechanical properties. It was our aim to evaluate if human adipoderived stem cells (ASCs) could replace tenocytes for scaffold seeding.

Methods:  ASCs and tenocytes were co-cultured in different ratios (3:1. 1:1, 1:3) and with three different methods (1. direct co-culture, 2. tenocyte-conditioned media on ASCs and 3. and an insert system to keep both cell types in same media without contact). Proliferation, collagen production and tenogenic marker expression were measured by hematocytometry, immunocytochemistry, ELISA and real-time PCR. 

Results: Proliferation and collagen production were similar for tenocytes and ASCs alone. Tenascin C was a reliable tenocyte marker. Proliferation was increased in direct co-culture, especially at an ASC:tenocyte ratio of 3:1, and for tenocytes in ASC-conditioned media. Direct co-culture caused significant upregulation in tenascin C expression in ASCs (4.0x, p<0.05). In tenocyte-conditioned media, tenascin C expression was upregulated 2.5x (p<0.05). In the insert system, tenascin C expression was upregulated 2.3x (p<0.05).

Conclusion: ASCs are good candidates for tendon tissue engineering because they are similar to tenocytes in proliferation and collagen production. They increase proliferation in co-culture and adapt a tenocyte-like phenotype. An ASC : tenocyte ratio of 3:1 is optimal to provide good proliferation and phenotype change by minimizing donor morbidity.

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