title: Isolation and trans-differentiation of mesenchymal stromal cells into smooth muscle cells: Utility and applicability for cell-sheet engineering.
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.
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA; Department of Cardiovascular Surgery, School of Medicine, Osaka University Graduate, Osaka, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shudo
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Jeffrey E
authors.initials: JE
authors.lastname: Cohen
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Goldstone
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: John W
authors.initials: JW
authors.lastname: MacArthur
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Jay
authors.initials: J
authors.lastname: Patel
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Bryan B
authors.initials: BB
authors.lastname: Edwards
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Hopkins
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Amanda N
authors.initials: AN
authors.lastname: Steele
authors.affiliation: Cell Sciences Imaging Facility, Stanford School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Lydia-Marie
authors.initials: LM
authors.lastname: Joubert
authors.affiliation: Department of Cardiovascular Surgery, School of Medicine, Osaka University Graduate, Osaka, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Miyagawa
authors.affiliation: Department of Cardiovascular Surgery, School of Medicine, Osaka University Graduate, Osaka, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sawa
authors.affiliation: Department of Cardiothoracic Surgery, School of Medicine, Stanford University, Stanford, CA, USA. Electronic address: joswoo@stanford.edu.
authors.forename: Y Joseph
authors.initials: YJ
authors.lastname: Woo
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D009130:Muscle, Smooth / Q000166:cytology / Q000502:physiology; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D016326:Extracellular Matrix Proteins
keywords: cell culture; co-culture; flow cytometry; scanning electron microscopy