Isolation and trans-differentiation of mesenchymal stromal cells into smooth muscle cells: Utility and applicability for cell-sheet engineering

• Published in: Cytotherapy (Oxford, England) England), 2016-04, Vol.18 (4), p.510-517
• Main Authors: Shudo, Yasuhiro, Cohen, Jeffrey E, Goldstone, Andrew B, MacArthur, John W, Patel, Jay, Edwards, Bryan B, Hopkins, Michael S, Steele, Amanda N, Joubert, Lydia-Marie, Miyagawa, Shigeru, Sawa, Yoshiki, Woo, Y. Joseph
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; Animals; cell culture; Cell Proliferation; Cell Separation - methods; Cell Transdifferentiation; Cells, Cultured; co-culture; Extracellular Matrix - metabolism; Extracellular Matrix Proteins - metabolism; flow cytometry; Male; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - physiology; Muscle, Smooth - cytology; Muscle, Smooth - physiology; Myocytes, Smooth Muscle - physiology; Other; Rats; Rats, Wistar; scanning electron microscopy; Tissue Engineering - methods
• ISSN: 1465-3249; 1477-2566
• DOI: 10.1016/j.jcyt.2016.01.012

Abstract Background Bone marrow (BM)-derived mesenchymal stromal cells (MSCs) have shown potential to differentiate into various cell types, including smooth muscle cells (SMCs). The extracellular matrix (ECM) represents an appealing and readily available source of SMCs for use in tissue engineering. In this study, we hypothesized that the ECM could be used to induce MSC differentiation to SMCs for engineered cell-sheet construction. Methods Primary MSCs were isolated from the BM of Wistar rats, transferred and cultured on dishes coated with 3 different types of ECM: collagen type IV (Col IV), fibronectin (FN), and laminin (LM). Primary MSCs were also included as a control. The proportions of SMC (a smooth muscle actin [aSMA] and SM22a) and MSC markers were examined with flow cytometry and Western blotting, and cell proliferation rates were also quantified. Results Both FN and LM groups were able to induce differentiation of MSCs toward smooth muscle–like cell types, as evidenced by an increase in the proportion of SMC markers (aSMA; Col IV 42.3 ± 6.9%, FN 65.1 ± 6.5%, LM 59.3 ± 7.0%, Control 39.9 ± 3.1%; P  = 0.02, SM22; Col IV 56.0 ± 7.7%, FN 74.2 ± 6.7%, LM 60.4 ± 8.7%, Control 44.9 ± 3.6%) and a decrease in that of MSC markers (CD105: Col IV 64.0 ± 5.2%, FN 57.6 ± 4.0%, LM 60.3 ± 7.0%, Control 85.3 ± 4.2%; P  = 0.03). The LM group showed a decrease in overall cell proliferation, whereas FN and Col IV groups remained similar to control MSCs (Col IV, 9.0 ± 2.3%; FN, 9.8 ± 2.5%; LM, 4.3 ± 1.3%; Control, 9.8 ± 2.8%). Conclusions Our findings indicate that ECM selection can guide differentiation of MSCs into the SMC lineage. Fibronectin preserved cellular proliferative capacity while yielding the highest proportion of differentiated SMCs, suggesting that FN-coated materials may be facilitate smooth muscle tissue engineering.