Factors affecting the structure and maturation of human tissue engineered skeletal muscle

Neil R W Martin, Samantha L. Passey, Darren J. Player, Alastair Khodabukus, Richard A. Ferguson, Adam P. Sharples, Vivek Mudera, Keith Baar, Mark P. Lewis

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Tissue engineered skeletal muscle has great utility in experimental studies of physiology, clinical testing and its potential for transplantation to replace damaged tissue. Despite recent work in rodent tissue or cell lines, there is a paucity of literature concerned with the culture of human muscle derived cells (MDCs) in engineered constructs. Here we aimed to tissue engineer for the first time in the literature human skeletal muscle in self-assembling fibrin hydrogels and determine the effect of MDC seeding density and myogenic proportion on the structure and maturation of the constructs. Constructs seeded with 4×105 MDCs assembled to a greater extent than those at 1×105 or 2×105, and immunostaining revealed a higher fusion index and a higher density of myotubes within the constructs, showing greater structural semblance to invivo tissue. These constructs primarily expressed perinatal and slow type I myosin heavy chain mRNA after 21 days in culture. In subsequent experiments MACS® technology was used to separate myogenic and non-myogenic cells from their heterogeneous parent population and these cells were seeded at varying myogenic (desmin+) proportions in fibrin based constructs. Only in the constructs seeded with 75% desmin+cells was there evidence of striations when immunostained for slow myosin heavy chain compared with constructs seeded with 10 or 50% desmin+cells. Overall, this work reveals the importance of cell number and myogenic proportions in tissue engineering human skeletal muscle with structural resemblance to invivo tissue.

Original languageEnglish (US)
Pages (from-to)5759-5765
Number of pages7
JournalBiomaterials
Volume34
Issue number23
DOIs
StatePublished - Jul 2013

Fingerprint

Muscle
Skeletal Muscle
Tissue
Desmin
Muscle Cells
Myosin Heavy Chains
Fibrin
Cell Count
Hydrogels
Skeletal Muscle Fibers
Physiology
Tissue Engineering
Tissue engineering
Rodentia
Fusion reactions
Transplantation
Cells
Technology
Engineers
Cell Line

Keywords

  • Human skeletal muscle
  • Muscle derived cells
  • Myosin heavy chain
  • Tissue engineering

ASJC Scopus subject areas

  • Biomaterials
  • Bioengineering
  • Ceramics and Composites
  • Mechanics of Materials
  • Biophysics

Cite this

Martin, N. R. W., Passey, S. L., Player, D. J., Khodabukus, A., Ferguson, R. A., Sharples, A. P., ... Lewis, M. P. (2013). Factors affecting the structure and maturation of human tissue engineered skeletal muscle. Biomaterials, 34(23), 5759-5765. https://doi.org/10.1016/j.biomaterials.2013.04.002

Factors affecting the structure and maturation of human tissue engineered skeletal muscle. / Martin, Neil R W; Passey, Samantha L.; Player, Darren J.; Khodabukus, Alastair; Ferguson, Richard A.; Sharples, Adam P.; Mudera, Vivek; Baar, Keith; Lewis, Mark P.

In: Biomaterials, Vol. 34, No. 23, 07.2013, p. 5759-5765.

Research output: Contribution to journalArticle

Martin, NRW, Passey, SL, Player, DJ, Khodabukus, A, Ferguson, RA, Sharples, AP, Mudera, V, Baar, K & Lewis, MP 2013, 'Factors affecting the structure and maturation of human tissue engineered skeletal muscle', Biomaterials, vol. 34, no. 23, pp. 5759-5765. https://doi.org/10.1016/j.biomaterials.2013.04.002
Martin NRW, Passey SL, Player DJ, Khodabukus A, Ferguson RA, Sharples AP et al. Factors affecting the structure and maturation of human tissue engineered skeletal muscle. Biomaterials. 2013 Jul;34(23):5759-5765. https://doi.org/10.1016/j.biomaterials.2013.04.002
Martin, Neil R W ; Passey, Samantha L. ; Player, Darren J. ; Khodabukus, Alastair ; Ferguson, Richard A. ; Sharples, Adam P. ; Mudera, Vivek ; Baar, Keith ; Lewis, Mark P. / Factors affecting the structure and maturation of human tissue engineered skeletal muscle. In: Biomaterials. 2013 ; Vol. 34, No. 23. pp. 5759-5765.
@article{7d7f511dff3748cea8f98657df710c06,
title = "Factors affecting the structure and maturation of human tissue engineered skeletal muscle",
abstract = "Tissue engineered skeletal muscle has great utility in experimental studies of physiology, clinical testing and its potential for transplantation to replace damaged tissue. Despite recent work in rodent tissue or cell lines, there is a paucity of literature concerned with the culture of human muscle derived cells (MDCs) in engineered constructs. Here we aimed to tissue engineer for the first time in the literature human skeletal muscle in self-assembling fibrin hydrogels and determine the effect of MDC seeding density and myogenic proportion on the structure and maturation of the constructs. Constructs seeded with 4×105 MDCs assembled to a greater extent than those at 1×105 or 2×105, and immunostaining revealed a higher fusion index and a higher density of myotubes within the constructs, showing greater structural semblance to invivo tissue. These constructs primarily expressed perinatal and slow type I myosin heavy chain mRNA after 21 days in culture. In subsequent experiments MACS{\circledR} technology was used to separate myogenic and non-myogenic cells from their heterogeneous parent population and these cells were seeded at varying myogenic (desmin+) proportions in fibrin based constructs. Only in the constructs seeded with 75{\%} desmin+cells was there evidence of striations when immunostained for slow myosin heavy chain compared with constructs seeded with 10 or 50{\%} desmin+cells. Overall, this work reveals the importance of cell number and myogenic proportions in tissue engineering human skeletal muscle with structural resemblance to invivo tissue.",
keywords = "Human skeletal muscle, Muscle derived cells, Myosin heavy chain, Tissue engineering",
author = "Martin, {Neil R W} and Passey, {Samantha L.} and Player, {Darren J.} and Alastair Khodabukus and Ferguson, {Richard A.} and Sharples, {Adam P.} and Vivek Mudera and Keith Baar and Lewis, {Mark P.}",
year = "2013",
month = "7",
doi = "10.1016/j.biomaterials.2013.04.002",
language = "English (US)",
volume = "34",
pages = "5759--5765",
journal = "Biomaterials",
issn = "0142-9612",
publisher = "Elsevier BV",
number = "23",

}

TY - JOUR

T1 - Factors affecting the structure and maturation of human tissue engineered skeletal muscle

AU - Martin, Neil R W

AU - Passey, Samantha L.

AU - Player, Darren J.

AU - Khodabukus, Alastair

AU - Ferguson, Richard A.

AU - Sharples, Adam P.

AU - Mudera, Vivek

AU - Baar, Keith

AU - Lewis, Mark P.

PY - 2013/7

Y1 - 2013/7

N2 - Tissue engineered skeletal muscle has great utility in experimental studies of physiology, clinical testing and its potential for transplantation to replace damaged tissue. Despite recent work in rodent tissue or cell lines, there is a paucity of literature concerned with the culture of human muscle derived cells (MDCs) in engineered constructs. Here we aimed to tissue engineer for the first time in the literature human skeletal muscle in self-assembling fibrin hydrogels and determine the effect of MDC seeding density and myogenic proportion on the structure and maturation of the constructs. Constructs seeded with 4×105 MDCs assembled to a greater extent than those at 1×105 or 2×105, and immunostaining revealed a higher fusion index and a higher density of myotubes within the constructs, showing greater structural semblance to invivo tissue. These constructs primarily expressed perinatal and slow type I myosin heavy chain mRNA after 21 days in culture. In subsequent experiments MACS® technology was used to separate myogenic and non-myogenic cells from their heterogeneous parent population and these cells were seeded at varying myogenic (desmin+) proportions in fibrin based constructs. Only in the constructs seeded with 75% desmin+cells was there evidence of striations when immunostained for slow myosin heavy chain compared with constructs seeded with 10 or 50% desmin+cells. Overall, this work reveals the importance of cell number and myogenic proportions in tissue engineering human skeletal muscle with structural resemblance to invivo tissue.

AB - Tissue engineered skeletal muscle has great utility in experimental studies of physiology, clinical testing and its potential for transplantation to replace damaged tissue. Despite recent work in rodent tissue or cell lines, there is a paucity of literature concerned with the culture of human muscle derived cells (MDCs) in engineered constructs. Here we aimed to tissue engineer for the first time in the literature human skeletal muscle in self-assembling fibrin hydrogels and determine the effect of MDC seeding density and myogenic proportion on the structure and maturation of the constructs. Constructs seeded with 4×105 MDCs assembled to a greater extent than those at 1×105 or 2×105, and immunostaining revealed a higher fusion index and a higher density of myotubes within the constructs, showing greater structural semblance to invivo tissue. These constructs primarily expressed perinatal and slow type I myosin heavy chain mRNA after 21 days in culture. In subsequent experiments MACS® technology was used to separate myogenic and non-myogenic cells from their heterogeneous parent population and these cells were seeded at varying myogenic (desmin+) proportions in fibrin based constructs. Only in the constructs seeded with 75% desmin+cells was there evidence of striations when immunostained for slow myosin heavy chain compared with constructs seeded with 10 or 50% desmin+cells. Overall, this work reveals the importance of cell number and myogenic proportions in tissue engineering human skeletal muscle with structural resemblance to invivo tissue.

KW - Human skeletal muscle

KW - Muscle derived cells

KW - Myosin heavy chain

KW - Tissue engineering

UR - http://www.scopus.com/inward/record.url?scp=84877837007&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84877837007&partnerID=8YFLogxK

U2 - 10.1016/j.biomaterials.2013.04.002

DO - 10.1016/j.biomaterials.2013.04.002

M3 - Article

C2 - 23643182

AN - SCOPUS:84877837007

VL - 34

SP - 5759

EP - 5765

JO - Biomaterials

JF - Biomaterials

SN - 0142-9612

IS - 23

ER -