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Table 3 Examples of tendon cell sheet models

From: Boosting tendon repair: interplay of cells, growth factors and scaffold-free and gel-based carriers

Experimental cell sheet Outcome Preparation of cell sheet Study type and animal models Reference
Rabbit ADSCs sheet Cell sheets were cultured over 3 weeks, and cell metabolic activity, cell sheet thickness, and early differentiation gene expression were analyzed. One week-old cell sheets displayed upregulation of early differentiation gene markers (Runx and Sox9). Cell sheet thickness and cell metabolic activity increased in the second and third week. ADSCs were cultured in 6-well culture plates until 100% confluence. Confluent cells were then cultured in expansion medium supplemented with 50 mg/ml ascorbic acid for 3 weeks to facilitate cell sheet formation. In vitro Neo et al. 2016
Mouse MSC sheet MSC sheet transplantation into musculotendinous junction at 4–8 weeks showed similar recovery of muscle mass and tension to the contralateral non-transplanted side. However, at 14–18 weeks, MSC sheet-treated group showed increased recovery of muscle mass and tension output. Engrafted MSCs primarily formed connective tissues and muscle fibres, and bridged the ruptured tendon-muscle fibre units. The cells reached full confluence, detached from culture dishes with 2 mM EDTA, then collected and centrifuged into hybrid sheet/pellet like structure. In vitro and in vivo; Mouse musculo-tendinous junction model Hashimoto et al. 2016
Human ACL-derived CD34+ cell sheet ACL-derived CD34+ cell sheet improved the ACL repair which was judged by histological assessment at week 2 and biomechanical evaluation at week 8 in a rat ACL injury model. Cells were plated in temperature-responsive culture dishes at 37 °C for 17 h, and then incubated at 20 °C for 20 min, and afterwards the cell sheets detached spontaneously. Rat ACL injury model Mifune et al. 2013
Human rotator cuff-derived cell sheet The cell sheets transplanted to the infraspinatus injury site induced angiogenesis and Col synthesis, and improved tendon-bone junction repair at 4 and 8 weeks postoperation. Cells were cultured on 24-well temperature-responsive culture dishes at 37 °C for 17 h. Then, the plates were placed at room temperature for 20 min, and the cell sheets detached from the wells spontaneously. Rat rotator cuff injury model Harada et al. 2017
Rat TSPC GFP-labelled sheet The TSPC sheet radiographically, histologically and biomechanically improved ACL healing in a rat model at week 2, 6 and 12 postoperatively. GFP-labelled TSPCs were detected at the graft-bone tunnel interface and in the intra-articular graft midsubstance in all samples at week 2. Cells were plated in normal culture dishes in low-glucose medium. After 100% confluence, cell sheet was detached by rinsing with saline. Rat ACL injury model Lui et al. 2014
Rat TSPC sheet TSPC sheet grafting into Achilles tendon defect significantly improved the histological features and Col content both at 2 and 4 weeks post-surgery, indicating that TSPC sheets can speed up tendon remodelling in the early stages of the healing process. TSPC sheets were prepared by plating on temperature-responsive culture dishes. Cells were cultured for 3 days and then induced for cell sheet formation by treating with 25 mM ascorbic acid in complete culture medium at 37 °C. After 9 days, monolayer cell sheets were obtained by reducing the temperature from 37 °C to 20 °C for 20 min. Rat Achilles tendon injury model Komatsu, et al. 2016