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Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue

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Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue. / Stoehr, Andrea; Hirt, Marc N; Hansen, Arne; Seiffert, Moritz; Conradi, Lenard; Uebeler, June; Limbourg, Florian P; Eschenhagen, Thomas.

In: TISSUE ENG PT A, Vol. 22, No. 3-4, 02.2016, p. 326-35.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Stoehr, A, Hirt, MN, Hansen, A, Seiffert, M, Conradi, L, Uebeler, J, Limbourg, FP & Eschenhagen, T 2016, 'Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue', TISSUE ENG PT A, vol. 22, no. 3-4, pp. 326-35. https://doi.org/10.1089/ten.TEA.2015.0242

APA

Stoehr, A., Hirt, M. N., Hansen, A., Seiffert, M., Conradi, L., Uebeler, J., Limbourg, F. P., & Eschenhagen, T. (2016). Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue. TISSUE ENG PT A, 22(3-4), 326-35. https://doi.org/10.1089/ten.TEA.2015.0242

Vancouver

Stoehr A, Hirt MN, Hansen A, Seiffert M, Conradi L, Uebeler J et al. Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue. TISSUE ENG PT A. 2016 Feb;22(3-4):326-35. https://doi.org/10.1089/ten.TEA.2015.0242

Bibtex

@article{b810a32d3e5a4fbe8eb849c62280415b,
title = "Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue",
abstract = "Engineered heart tissue (EHT) from primary heart cells contains endothelial cells (ECs), but the extent to which ECs organize into vessel-like structures or even functional vessels remains unknown and is difficult to study by conventional methods. In this study, we generated fibrin-based mini-EHTs from a transgenic mouse line (Cdh5-CreERT2 × Rosa26-LacZ), in which ECs were specifically and inducibly labeled by applying tamoxifen (EC(iLacZ)). EHTs were generated from an unpurified cell mix of newborn mouse hearts and were cultured under standard serum-containing conditions. Cre expression in 15-day-old EHTs was induced by addition of o-hydroxytamoxifen to the culture medium for 48 h, and ECs were visualized by X-gal staining. EC(iLacZ) EHTs showed a dense X-gal-positive vessel-like network with distinct tubular structures. Immunofluorescence revealed that ECs were mainly associated with cardiomyocytes within the EHT. EC(iLacZ) EHT developed spontaneous and regular contractility with forces up to 0.1 mN. Coherent contractility and the presence of an extensive vessel-like network were both dependent on the presence of animal sera in the culture medium. Contractile EC(iLacZ) EHTs successfully served as grafts in implantation studies onto the hearts of immunodeficient mice. Four weeks after implantation, EHTs showed X-gal-positive lumen-forming vessel structures connected to the host myocardium circulation as they contained erythrocytes on a regular basis. Taken together, genetic labeling of ECs revealed the extensive formation of vessel-like structures in EHTs in vitro. The EC(iLacZ) EHT model could help simultaneously study biological effects of compounds on cardiomyocyte function and tissue vascularization.",
keywords = "Animals, Coronary Vessels, Mice, Mice, SCID, Mice, Transgenic, Myocardium, Myocytes, Cardiac, Neovascularization, Physiologic, Tissue Engineering, Journal Article, Research Support, Non-U.S. Gov't",
author = "Andrea Stoehr and Hirt, {Marc N} and Arne Hansen and Moritz Seiffert and Lenard Conradi and June Uebeler and Limbourg, {Florian P} and Thomas Eschenhagen",
year = "2016",
month = feb,
doi = "10.1089/ten.TEA.2015.0242",
language = "English",
volume = "22",
pages = "326--35",
journal = "TISSUE ENG PT A",
issn = "1937-3341",
publisher = "Mary Ann Liebert Inc.",
number = "3-4",

}

RIS

TY - JOUR

T1 - Spontaneous Formation of Extensive Vessel-Like Structures in Murine Engineered Heart Tissue

AU - Stoehr, Andrea

AU - Hirt, Marc N

AU - Hansen, Arne

AU - Seiffert, Moritz

AU - Conradi, Lenard

AU - Uebeler, June

AU - Limbourg, Florian P

AU - Eschenhagen, Thomas

PY - 2016/2

Y1 - 2016/2

N2 - Engineered heart tissue (EHT) from primary heart cells contains endothelial cells (ECs), but the extent to which ECs organize into vessel-like structures or even functional vessels remains unknown and is difficult to study by conventional methods. In this study, we generated fibrin-based mini-EHTs from a transgenic mouse line (Cdh5-CreERT2 × Rosa26-LacZ), in which ECs were specifically and inducibly labeled by applying tamoxifen (EC(iLacZ)). EHTs were generated from an unpurified cell mix of newborn mouse hearts and were cultured under standard serum-containing conditions. Cre expression in 15-day-old EHTs was induced by addition of o-hydroxytamoxifen to the culture medium for 48 h, and ECs were visualized by X-gal staining. EC(iLacZ) EHTs showed a dense X-gal-positive vessel-like network with distinct tubular structures. Immunofluorescence revealed that ECs were mainly associated with cardiomyocytes within the EHT. EC(iLacZ) EHT developed spontaneous and regular contractility with forces up to 0.1 mN. Coherent contractility and the presence of an extensive vessel-like network were both dependent on the presence of animal sera in the culture medium. Contractile EC(iLacZ) EHTs successfully served as grafts in implantation studies onto the hearts of immunodeficient mice. Four weeks after implantation, EHTs showed X-gal-positive lumen-forming vessel structures connected to the host myocardium circulation as they contained erythrocytes on a regular basis. Taken together, genetic labeling of ECs revealed the extensive formation of vessel-like structures in EHTs in vitro. The EC(iLacZ) EHT model could help simultaneously study biological effects of compounds on cardiomyocyte function and tissue vascularization.

AB - Engineered heart tissue (EHT) from primary heart cells contains endothelial cells (ECs), but the extent to which ECs organize into vessel-like structures or even functional vessels remains unknown and is difficult to study by conventional methods. In this study, we generated fibrin-based mini-EHTs from a transgenic mouse line (Cdh5-CreERT2 × Rosa26-LacZ), in which ECs were specifically and inducibly labeled by applying tamoxifen (EC(iLacZ)). EHTs were generated from an unpurified cell mix of newborn mouse hearts and were cultured under standard serum-containing conditions. Cre expression in 15-day-old EHTs was induced by addition of o-hydroxytamoxifen to the culture medium for 48 h, and ECs were visualized by X-gal staining. EC(iLacZ) EHTs showed a dense X-gal-positive vessel-like network with distinct tubular structures. Immunofluorescence revealed that ECs were mainly associated with cardiomyocytes within the EHT. EC(iLacZ) EHT developed spontaneous and regular contractility with forces up to 0.1 mN. Coherent contractility and the presence of an extensive vessel-like network were both dependent on the presence of animal sera in the culture medium. Contractile EC(iLacZ) EHTs successfully served as grafts in implantation studies onto the hearts of immunodeficient mice. Four weeks after implantation, EHTs showed X-gal-positive lumen-forming vessel structures connected to the host myocardium circulation as they contained erythrocytes on a regular basis. Taken together, genetic labeling of ECs revealed the extensive formation of vessel-like structures in EHTs in vitro. The EC(iLacZ) EHT model could help simultaneously study biological effects of compounds on cardiomyocyte function and tissue vascularization.

KW - Animals

KW - Coronary Vessels

KW - Mice

KW - Mice, SCID

KW - Mice, Transgenic

KW - Myocardium

KW - Myocytes, Cardiac

KW - Neovascularization, Physiologic

KW - Tissue Engineering

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1089/ten.TEA.2015.0242

DO - 10.1089/ten.TEA.2015.0242

M3 - SCORING: Journal article

C2 - 26763667

VL - 22

SP - 326

EP - 335

JO - TISSUE ENG PT A

JF - TISSUE ENG PT A

SN - 1937-3341

IS - 3-4

ER -