TY - JOUR
T1 - Developing 3D Organized Human Cardiac Tissue within a Microfluidic Platform
AU - Veldhuizen, Jaimeson
AU - Nikkhah, Mehdi
N1 - Funding Information:
We would like to thank NSF CAREER Award #1653193, Arizona Biomedical Research Commission (ABRC) New Investigator Award (ADHS18-198872), and the Flinn Foundation Award for providing funding sources for this project. The hiPSC line, SCVI20, was obtained from Joseph C. Wu, MD, PhD at the Stanford Cardiovascular Institute funded by NIH R24 HL117756. The hiPSC line, IMR90-4, was obtained from WiCell Research Institute55,56.
Funding Information:
We would like to thank NSF CAREER Award #1653193, Arizona Biomedical Research Commission (ABRC) New Investigator Award (ADHS18-198872), and the Flinn Foundation Award for providing funding sources for this project. The hiPSC line, SCVI20, was obtained from Joseph C. Wu, MD, PhD at the Stanford Cardiovascular Institute funded by NIH R24 HL117756. The hiPSC line, IMR90-4, was obtained from WiCell Research Institute55,56 .
Publisher Copyright:
© 2021 JoVE Journal of Visualized Experiments.
PY - 2021/5
Y1 - 2021/5
N2 - The leading cause of death worldwide persists as cardiovascular disease (CVD). However, modeling the physiological and biological complexity of the heart muscle, the myocardium, is notoriously difficult to accomplish in vitro. Mainly, obstacles lie in the need for human cardiomyocytes (CMs) that are either adult or exhibit adult-like phenotypes and can successfully replicate the myocardium's cellular complexity and intricate 3D architecture. Unfortunately, due to ethical concerns and lack of available primary patient-derived human cardiac tissue, combined with the minimal proliferation of CMs, the sourcing of viable human CMs has been a limiting step for cardiac tissue engineering. To this end, most research has transitioned toward cardiac differentiation of human induced pluripotent stem cells (hiPSCs) as the primary source of human CMs, resulting in the wide incorporation of hiPSC-CMs within in vitro assays for cardiac tissue modeling. Here in this work, we demonstrate a protocol for developing a 3D mature stem cell-derived human cardiac tissue within a microfluidic device. We specifically explain and visually demonstrate the production of a 3D in vitro anisotropic cardiac tissue-on-a-chip model from hiPSC-derived CMs. We primarily describe a purification protocol to select for CMs, the co-culture of cells with a defined ratio via mixing CMs with human CFs (hCFs), and suspension of this co-culture within the collagen-based hydrogel. We further demonstrate the injection of the cell-laden hydrogel within our well-defined microfluidic device, embedded with staggered elliptical microposts that serve as surface topography to induce a high degree of alignment of the surrounding cells and the hydrogel matrix, mimicking the architecture of the native myocardium. We envision that the proposed 3D anisotropic cardiac tissue-on-chip model is suitable for fundamental biology studies, disease modeling, and, through its use as a screening tool, pharmaceutical testing.
AB - The leading cause of death worldwide persists as cardiovascular disease (CVD). However, modeling the physiological and biological complexity of the heart muscle, the myocardium, is notoriously difficult to accomplish in vitro. Mainly, obstacles lie in the need for human cardiomyocytes (CMs) that are either adult or exhibit adult-like phenotypes and can successfully replicate the myocardium's cellular complexity and intricate 3D architecture. Unfortunately, due to ethical concerns and lack of available primary patient-derived human cardiac tissue, combined with the minimal proliferation of CMs, the sourcing of viable human CMs has been a limiting step for cardiac tissue engineering. To this end, most research has transitioned toward cardiac differentiation of human induced pluripotent stem cells (hiPSCs) as the primary source of human CMs, resulting in the wide incorporation of hiPSC-CMs within in vitro assays for cardiac tissue modeling. Here in this work, we demonstrate a protocol for developing a 3D mature stem cell-derived human cardiac tissue within a microfluidic device. We specifically explain and visually demonstrate the production of a 3D in vitro anisotropic cardiac tissue-on-a-chip model from hiPSC-derived CMs. We primarily describe a purification protocol to select for CMs, the co-culture of cells with a defined ratio via mixing CMs with human CFs (hCFs), and suspension of this co-culture within the collagen-based hydrogel. We further demonstrate the injection of the cell-laden hydrogel within our well-defined microfluidic device, embedded with staggered elliptical microposts that serve as surface topography to induce a high degree of alignment of the surrounding cells and the hydrogel matrix, mimicking the architecture of the native myocardium. We envision that the proposed 3D anisotropic cardiac tissue-on-chip model is suitable for fundamental biology studies, disease modeling, and, through its use as a screening tool, pharmaceutical testing.
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UR - http://www.scopus.com/inward/citedby.url?scp=85110827797&partnerID=8YFLogxK
U2 - 10.3791/62539
DO - 10.3791/62539
M3 - Article
C2 - 34223835
AN - SCOPUS:85110827797
VL - 2021
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
SN - 1940-087X
IS - 172
M1 - e62539
ER -