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A three-dimensional gel bioreactor for assessment of cardiomyocyte induction in skeletal muscle-derived stem cells

Clause, KC and Tinney, JP and Liu, LJ and Gharaibeh, B and Huard, J and Kirk, JA and Shroff, SG and Fujimoto, KL and Wagner, WR and Ralphe, JC and Keller, BB and Tobita, K (2010) A three-dimensional gel bioreactor for assessment of cardiomyocyte induction in skeletal muscle-derived stem cells. Tissue Engineering - Part C: Methods, 16 (3). 375 - 385. ISSN 1937-3384

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Abstract

Skeletal muscle-derived stem cells (MDSCs) are able to differentiate into cardiomyocytes (CMs). However, it remains to be investigated whether differentiated CMs contract similar to native CMs. Here, we developed a three-dimensional collagen gel bioreactor (3DGB) that induces a working CM phenotype from MDSCs, and the contractile properties are directly measured as an engineered cardiac tissue. Neonate rat MDSCs were isolated from hind-leg muscles via the preplate technique. Isolated MDSCs were approximately 60% positive to Sca-1 and negative to CD34, CD45, or c-kit antigens. We sorted Sca-1(-) MDSCs and constructed MDSC-3DGBs by mixing MDSCs with acid soluble rat tail collagen type-I and matrix factors. MDSC-3DGB exhibited spontaneous cyclic contraction by culture day 7. MDSC-3DGB expressed cardiac-specific genes and proteins. Histological assessment revealed that cardiac-specific troponin-T and-I expressed in a typical striation pattern and connexin-43 was expressed similar to the native fetal ventricular papillary muscle. β-Adrenergic stimulation increased MDSC-3DGB spontaneous beat frequency. MDSC-3DGB generated contractile force and intracellular calcium ion transients similar to engineered cardiac tissue from native cardiac cells. Results suggest that MDSC-3DGB induces a working CM phenotype in MDSCs and is a useful 3D culture system to directly assess the contractile properties of differentiated CMs in vitro. © 2010 Mary Ann Liebert, Inc.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Clause, KC
Tinney, JP
Liu, LJljl4@pitt.eduLJL4
Gharaibeh, Bburhan@pitt.eduBURHAN0000-0002-5947-1232
Huard, J
Kirk, JA
Shroff, SGsshroff@pitt.eduSSHROFF
Fujimoto, KL
Wagner, WRwagner@pitt.eduWAGNER0000-0003-0082-8089
Ralphe, JC
Keller, BB
Tobita, Kkit3@pitt.eduKIT3
Date: 1 June 2010
Date Type: Publication
Journal or Publication Title: Tissue Engineering - Part C: Methods
Volume: 16
Number: 3
Page Range: 375 - 385
DOI or Unique Handle: 10.1089/ten.tec.2009.0098
Schools and Programs: School of Medicine > Orthopaedic Surgery
School of Medicine > Pediatrics
School of Medicine > Surgery
Swanson School of Engineering > Bioengineering
Refereed: Yes
ISSN: 1937-3384
MeSH Headings: Animals; Animals, Newborn; Base Sequence; Bioreactors; Blotting, Western; Calcium--metabolism; Cell Differentiation; Cells, Cultured; Collagen; DNA Primers; Electrophoresis, Polyacrylamide Gel; Muscle, Skeletal--cytology; Muscle, Skeletal--metabolism; Myocardium--cytology; Myocardium--metabolism; Polymerase Chain Reaction; Rats; Rats, Inbred Lew; Stem Cells--cytology
Other ID: NLM PMC2945363
PubMed Central ID: PMC2945363
PubMed ID: 19601695
Date Deposited: 04 Apr 2014 16:41
Last Modified: 19 Mar 2024 19:55
URI: http://d-scholarship.pitt.edu/id/eprint/20948

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