Cargando…
Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle
Vascularization of tissue-engineered constructs remains a key challenge in the field of skeletal muscle tissue engineering. One strategy for vascularizing organoids is in vitro pre-vascularization, relying on de novo assembly of undifferentiated endothelial cells into capillaries, a process termed v...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9534561/ https://www.ncbi.nlm.nih.gov/pubmed/36213083 http://dx.doi.org/10.3389/fbioe.2022.964705 |
_version_ | 1784802570223484928 |
---|---|
author | Wüst, Rebecca Terrie, Lisanne Müntefering, Thomas Ruck, Tobias Thorrez, Lieven |
author_facet | Wüst, Rebecca Terrie, Lisanne Müntefering, Thomas Ruck, Tobias Thorrez, Lieven |
author_sort | Wüst, Rebecca |
collection | PubMed |
description | Vascularization of tissue-engineered constructs remains a key challenge in the field of skeletal muscle tissue engineering. One strategy for vascularizing organoids is in vitro pre-vascularization, relying on de novo assembly of undifferentiated endothelial cells into capillaries, a process termed vasculogenesis. In most endothelial cell research to date, human umbilical vein endothelial cells have been used primarily because of their availability. Nevertheless, this endothelial cell type is naturally not occurring in skeletal muscle tissue. Since endothelial cells display a tissue-specific phenotype, it is of interest to use muscle-specific microvascular endothelial cells to study pre-vascularization in skeletal muscle tissue engineering research. Thus far, tissue biopsies had to be processed in two separate protocols to obtain cells from the myogenic and the endothelial compartment. Here, we describe a novel, detailed protocol for the co-isolation of human skeletal muscle microvascular endothelial cells and satellite cell-derived myoblasts. It incorporates an automated mechanical and enzymatic tissue dissociation followed by magnetically activated cell sorting based on a combination of endothelial and skeletal muscle cell markers. Qualitative, quantitative, and functional characterization of the obtained cells is described and demonstrated by representative results. The simultaneous isolation of both cell types from the same donor is advantageous in terms of time efficiency. In addition, it may be the only possible method to isolate both cell types as the amount of tissue biopsy is often limited. The isolation of the two cell types is crucial for further studies to elucidate cell crosstalk in health and disease. Furthermore, the use of muscle-specific microvascular endothelial cells allows a shift towards engineering more physiologically relevant functional tissue, with downstream applications including drug screening and regenerative medicine. |
format | Online Article Text |
id | pubmed-9534561 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95345612022-10-06 Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle Wüst, Rebecca Terrie, Lisanne Müntefering, Thomas Ruck, Tobias Thorrez, Lieven Front Bioeng Biotechnol Bioengineering and Biotechnology Vascularization of tissue-engineered constructs remains a key challenge in the field of skeletal muscle tissue engineering. One strategy for vascularizing organoids is in vitro pre-vascularization, relying on de novo assembly of undifferentiated endothelial cells into capillaries, a process termed vasculogenesis. In most endothelial cell research to date, human umbilical vein endothelial cells have been used primarily because of their availability. Nevertheless, this endothelial cell type is naturally not occurring in skeletal muscle tissue. Since endothelial cells display a tissue-specific phenotype, it is of interest to use muscle-specific microvascular endothelial cells to study pre-vascularization in skeletal muscle tissue engineering research. Thus far, tissue biopsies had to be processed in two separate protocols to obtain cells from the myogenic and the endothelial compartment. Here, we describe a novel, detailed protocol for the co-isolation of human skeletal muscle microvascular endothelial cells and satellite cell-derived myoblasts. It incorporates an automated mechanical and enzymatic tissue dissociation followed by magnetically activated cell sorting based on a combination of endothelial and skeletal muscle cell markers. Qualitative, quantitative, and functional characterization of the obtained cells is described and demonstrated by representative results. The simultaneous isolation of both cell types from the same donor is advantageous in terms of time efficiency. In addition, it may be the only possible method to isolate both cell types as the amount of tissue biopsy is often limited. The isolation of the two cell types is crucial for further studies to elucidate cell crosstalk in health and disease. Furthermore, the use of muscle-specific microvascular endothelial cells allows a shift towards engineering more physiologically relevant functional tissue, with downstream applications including drug screening and regenerative medicine. Frontiers Media S.A. 2022-09-21 /pmc/articles/PMC9534561/ /pubmed/36213083 http://dx.doi.org/10.3389/fbioe.2022.964705 Text en Copyright © 2022 Wüst, Terrie, Müntefering, Ruck and Thorrez. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Wüst, Rebecca Terrie, Lisanne Müntefering, Thomas Ruck, Tobias Thorrez, Lieven Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
title | Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
title_full | Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
title_fullStr | Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
title_full_unstemmed | Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
title_short | Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
title_sort | efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9534561/ https://www.ncbi.nlm.nih.gov/pubmed/36213083 http://dx.doi.org/10.3389/fbioe.2022.964705 |
work_keys_str_mv | AT wustrebecca efficientcoisolationofmicrovascularendothelialcellsandsatellitecellderivedmyoblastsfromhumanskeletalmuscle AT terrielisanne efficientcoisolationofmicrovascularendothelialcellsandsatellitecellderivedmyoblastsfromhumanskeletalmuscle AT munteferingthomas efficientcoisolationofmicrovascularendothelialcellsandsatellitecellderivedmyoblastsfromhumanskeletalmuscle AT rucktobias efficientcoisolationofmicrovascularendothelialcellsandsatellitecellderivedmyoblastsfromhumanskeletalmuscle AT thorrezlieven efficientcoisolationofmicrovascularendothelialcellsandsatellitecellderivedmyoblastsfromhumanskeletalmuscle |