Abstract

Development of an ex vivo culture system to expand satellite cells, the resident muscle stem cell population, will be necessary for the development of their use as therapeutics. The loss of the niche environment is often cited as the reason that culture results in both the loss of myogenic potential and low re-engraftment rates of these cells. Studies have shown that culture of satellite cells on more elastic substrates maintained their quiescence and potential and increased re-engraftment, but there was limited proliferation. We examined whether substrates composed of extracellular matrix proteins, as either coatings or hydrogels, could support expansion of this population whilst maintaining the potency of these cells. The collagen based hydrogels were very pliant and our studies demonstrate that stiffer substrates are necessary for in vitro proliferation and differentiation of satellite cells, and the ECM composition was not significantly important. Our data further indicates that culture on highly elastic substrates allowed satellite cells to down-regulate myogenic specific transcription factors, resulting in an expression profile similar to a Galert state. These satellite cells could be subsequently cultured on Matrigel and induced to differentiate. Proliferation and gene expression data further indicated that C2C12 cells are not a good proxy for studies of satellite cell proliferation and differentiation on alternative substrates. Statement of Significance: Although biomaterials and muscle stem cell-based therapeutics for muscle loss due to trauma and disease have been studied intensely for the last decade, significant challenges remain; satellite cells lose their myogenic potential after in vitro culture, and do not re-engraft efficiently when delivered to skeletal muscle. We cultured adult mouse derived satellite cells and C2C12 cells on biomimetic, extracellular matrix based, hydrogels and coated plates and carried out a detailed analysis of proliferation, differentiation, and gene expression. These studies were designed to allow for the examination of the roles of both matrix composition and elasticity. The data demonstrated that satellite cells were most affected by elasticity. Examining the expression of lineage and cell cycle specific genes provided important insight into the behavior of both cell types, and point to fundamental differences that affect the interpretation of studies aimed at understanding the in vitro requirements of muscle progenitor cells. Understanding how satellite cells respond to various biochemical and biophysical cues at the molecular level will inform future efforts to design therapeutics for muscle regeneration.

Original languageEnglish (US)
JournalActa Biomaterialia
DOIs
StateAccepted/In press - Jan 1 2019

Fingerprint

Hydrogels
Muscle Cells
Extracellular Matrix
Muscle
Stem Cells
Satellites
Coatings
Substrates
Stem cells
Gene expression
Elasticity
Cell Differentiation
Military electronic countermeasures
Transcription factors
Extracellular Matrix Proteins
Cell proliferation
Gene Expression
Biomimetics
Biocompatible Materials
cdc Genes

Keywords

  • Extracellular matrix
  • Hydrogel
  • Muscle
  • Regeneration
  • Satellite cells

ASJC Scopus subject areas

  • Biotechnology
  • Biomaterials
  • Biochemistry
  • Biomedical Engineering
  • Molecular Biology

Cite this

@article{54d5d2dceae7400f8ff418f06b4d29a3,
title = "Molecular analysis of muscle progenitor cells on extracellular matrix coatings and hydrogels",
abstract = "Development of an ex vivo culture system to expand satellite cells, the resident muscle stem cell population, will be necessary for the development of their use as therapeutics. The loss of the niche environment is often cited as the reason that culture results in both the loss of myogenic potential and low re-engraftment rates of these cells. Studies have shown that culture of satellite cells on more elastic substrates maintained their quiescence and potential and increased re-engraftment, but there was limited proliferation. We examined whether substrates composed of extracellular matrix proteins, as either coatings or hydrogels, could support expansion of this population whilst maintaining the potency of these cells. The collagen based hydrogels were very pliant and our studies demonstrate that stiffer substrates are necessary for in vitro proliferation and differentiation of satellite cells, and the ECM composition was not significantly important. Our data further indicates that culture on highly elastic substrates allowed satellite cells to down-regulate myogenic specific transcription factors, resulting in an expression profile similar to a Galert state. These satellite cells could be subsequently cultured on Matrigel and induced to differentiate. Proliferation and gene expression data further indicated that C2C12 cells are not a good proxy for studies of satellite cell proliferation and differentiation on alternative substrates. Statement of Significance: Although biomaterials and muscle stem cell-based therapeutics for muscle loss due to trauma and disease have been studied intensely for the last decade, significant challenges remain; satellite cells lose their myogenic potential after in vitro culture, and do not re-engraft efficiently when delivered to skeletal muscle. We cultured adult mouse derived satellite cells and C2C12 cells on biomimetic, extracellular matrix based, hydrogels and coated plates and carried out a detailed analysis of proliferation, differentiation, and gene expression. These studies were designed to allow for the examination of the roles of both matrix composition and elasticity. The data demonstrated that satellite cells were most affected by elasticity. Examining the expression of lineage and cell cycle specific genes provided important insight into the behavior of both cell types, and point to fundamental differences that affect the interpretation of studies aimed at understanding the in vitro requirements of muscle progenitor cells. Understanding how satellite cells respond to various biochemical and biophysical cues at the molecular level will inform future efforts to design therapeutics for muscle regeneration.",
keywords = "Extracellular matrix, Hydrogel, Muscle, Regeneration, Satellite cells",
author = "Joanna Palade and Amrita Pal and Jeffery Rawls and Sarah Stabenfeldt and Norma Wilson-Rawls",
year = "2019",
month = "1",
day = "1",
doi = "10.1016/j.actbio.2019.08.019",
language = "English (US)",
journal = "Acta Biomaterialia",
issn = "1742-7061",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Molecular analysis of muscle progenitor cells on extracellular matrix coatings and hydrogels

AU - Palade, Joanna

AU - Pal, Amrita

AU - Rawls, Jeffery

AU - Stabenfeldt, Sarah

AU - Wilson-Rawls, Norma

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Development of an ex vivo culture system to expand satellite cells, the resident muscle stem cell population, will be necessary for the development of their use as therapeutics. The loss of the niche environment is often cited as the reason that culture results in both the loss of myogenic potential and low re-engraftment rates of these cells. Studies have shown that culture of satellite cells on more elastic substrates maintained their quiescence and potential and increased re-engraftment, but there was limited proliferation. We examined whether substrates composed of extracellular matrix proteins, as either coatings or hydrogels, could support expansion of this population whilst maintaining the potency of these cells. The collagen based hydrogels were very pliant and our studies demonstrate that stiffer substrates are necessary for in vitro proliferation and differentiation of satellite cells, and the ECM composition was not significantly important. Our data further indicates that culture on highly elastic substrates allowed satellite cells to down-regulate myogenic specific transcription factors, resulting in an expression profile similar to a Galert state. These satellite cells could be subsequently cultured on Matrigel and induced to differentiate. Proliferation and gene expression data further indicated that C2C12 cells are not a good proxy for studies of satellite cell proliferation and differentiation on alternative substrates. Statement of Significance: Although biomaterials and muscle stem cell-based therapeutics for muscle loss due to trauma and disease have been studied intensely for the last decade, significant challenges remain; satellite cells lose their myogenic potential after in vitro culture, and do not re-engraft efficiently when delivered to skeletal muscle. We cultured adult mouse derived satellite cells and C2C12 cells on biomimetic, extracellular matrix based, hydrogels and coated plates and carried out a detailed analysis of proliferation, differentiation, and gene expression. These studies were designed to allow for the examination of the roles of both matrix composition and elasticity. The data demonstrated that satellite cells were most affected by elasticity. Examining the expression of lineage and cell cycle specific genes provided important insight into the behavior of both cell types, and point to fundamental differences that affect the interpretation of studies aimed at understanding the in vitro requirements of muscle progenitor cells. Understanding how satellite cells respond to various biochemical and biophysical cues at the molecular level will inform future efforts to design therapeutics for muscle regeneration.

AB - Development of an ex vivo culture system to expand satellite cells, the resident muscle stem cell population, will be necessary for the development of their use as therapeutics. The loss of the niche environment is often cited as the reason that culture results in both the loss of myogenic potential and low re-engraftment rates of these cells. Studies have shown that culture of satellite cells on more elastic substrates maintained their quiescence and potential and increased re-engraftment, but there was limited proliferation. We examined whether substrates composed of extracellular matrix proteins, as either coatings or hydrogels, could support expansion of this population whilst maintaining the potency of these cells. The collagen based hydrogels were very pliant and our studies demonstrate that stiffer substrates are necessary for in vitro proliferation and differentiation of satellite cells, and the ECM composition was not significantly important. Our data further indicates that culture on highly elastic substrates allowed satellite cells to down-regulate myogenic specific transcription factors, resulting in an expression profile similar to a Galert state. These satellite cells could be subsequently cultured on Matrigel and induced to differentiate. Proliferation and gene expression data further indicated that C2C12 cells are not a good proxy for studies of satellite cell proliferation and differentiation on alternative substrates. Statement of Significance: Although biomaterials and muscle stem cell-based therapeutics for muscle loss due to trauma and disease have been studied intensely for the last decade, significant challenges remain; satellite cells lose their myogenic potential after in vitro culture, and do not re-engraft efficiently when delivered to skeletal muscle. We cultured adult mouse derived satellite cells and C2C12 cells on biomimetic, extracellular matrix based, hydrogels and coated plates and carried out a detailed analysis of proliferation, differentiation, and gene expression. These studies were designed to allow for the examination of the roles of both matrix composition and elasticity. The data demonstrated that satellite cells were most affected by elasticity. Examining the expression of lineage and cell cycle specific genes provided important insight into the behavior of both cell types, and point to fundamental differences that affect the interpretation of studies aimed at understanding the in vitro requirements of muscle progenitor cells. Understanding how satellite cells respond to various biochemical and biophysical cues at the molecular level will inform future efforts to design therapeutics for muscle regeneration.

KW - Extracellular matrix

KW - Hydrogel

KW - Muscle

KW - Regeneration

KW - Satellite cells

UR - http://www.scopus.com/inward/record.url?scp=85071333154&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85071333154&partnerID=8YFLogxK

U2 - 10.1016/j.actbio.2019.08.019

DO - 10.1016/j.actbio.2019.08.019

M3 - Article

JO - Acta Biomaterialia

JF - Acta Biomaterialia

SN - 1742-7061

ER -