Molly E Kupfer1,2, Wei-Han Lin1,2, Vasanth Ravikumar3, Kaiyan Qiu4, Lu Wang5, Ling Gao5, Didarul B Bhuiyan1, Megan Lenz1, Jeffrey Ai1, Ryan R Mahutga1, DeWayne Townsend6,7, Jianyi Zhang5, Michael C McAlpine4, Elena G Tolkacheva1,6,8, Brenda M Ogle1,2,6,8,9. 1. From the Department of Biomedical Engineering (M.E.K., W.-H.L., D.B.B., M.L., J.A., R.R.M., E.G.T., B.M.O.), University of Minnesota-Twin Cities, Minneapolis. 2. Stem Cell Institute (M.E.K., W.-H.L., B.M.O.), University of Minnesota-Twin Cities, Minneapolis. 3. Department of Electrical Engineering (V.R.), University of Minnesota-Twin Cities, Minneapolis. 4. Department of Mechanical Engineering (K.Q., M.C.M.), University of Minnesota-Twin Cities, Minneapolis. 5. Department of Biomedical Engineering, School of Medicine, School of Engineering, University of Alabama at Birmingham (L.W., L.G., J.Z.). 6. Lillehei Heart Institute (D.T., E.G.T., B.M.O.), University of Minnesota-Twin Cities, Minneapolis. 7. Department of Integrative Biology and Physiology (D.T.), University of Minnesota-Twin Cities, Minneapolis. 8. Institute for Engineering in Medicine (E.G.T., B.M.O.), University of Minnesota-Twin Cities, Minneapolis. 9. Masonic Cancer Center (B.M.O.), University of Minnesota-Twin Cities, Minneapolis.
Abstract
RATIONALE: One goal of cardiac tissue engineering is the generation of a living, human pump in vitro that could replace animal models and eventually serve as an in vivo therapeutic. Models that replicate the geometrically complex structure of the heart, harboring chambers and large vessels with soft biomaterials, can be achieved using 3-dimensional bioprinting. Yet, inclusion of contiguous, living muscle to support pump function has not been achieved. This is largely due to the challenge of attaining high densities of cardiomyocytes-a notoriously nonproliferative cell type. An alternative strategy is to print with human induced pluripotent stem cells, which can proliferate to high densities and fill tissue spaces, and subsequently differentiate them into cardiomyocytes in situ. OBJECTIVE: To develop a bioink capable of promoting human induced pluripotent stem cell proliferation and cardiomyocyte differentiation to 3-dimensionally print electromechanically functional, chambered organoids composed of contiguous cardiac muscle. METHODS AND RESULTS: We optimized a photo-crosslinkable formulation of native ECM (extracellular matrix) proteins and used this bioink to 3-dimensionally print human induced pluripotent stem cell-laden structures with 2 chambers and a vessel inlet and outlet. After human induced pluripotent stem cells proliferated to a sufficient density, we differentiated the cells within the structure and demonstrated function of the resultant human chambered muscle pump. Human chambered muscle pumps demonstrated macroscale beating and continuous action potential propagation with responsiveness to drugs and pacing. The connected chambers allowed for perfusion and enabled replication of pressure/volume relationships fundamental to the study of heart function and remodeling with health and disease. CONCLUSIONS: This advance represents a critical step toward generating macroscale tissues, akin to aggregate-based organoids, but with the critical advantage of harboring geometric structures essential to the pump function of cardiac muscle. Looking forward, human chambered organoids of this type might also serve as a test bed for cardiac medical devices and eventually lead to therapeutic tissue grafting.
RATIONALE: One goal of cardiac tissue engineering is the generation of a living, human pump in vitro that could replace animal models and eventually serve as an in vivo therapeutic. Models that replicate the geometrically complex structure of the heart, harboring chambers and large vessels with soft biomaterials, can be achieved using 3-dimensional bioprinting. Yet, inclusion of contiguous, living muscle to support pump function has not been achieved. This is largely due to the challenge of attaining high densities of cardiomyocytes-a notoriously nonproliferative cell type. An alternative strategy is to print with human induced pluripotent stem cells, which can proliferate to high densities and fill tissue spaces, and subsequently differentiate them into cardiomyocytes in situ. OBJECTIVE: To develop a bioink capable of promoting human induced pluripotent stem cell proliferation and cardiomyocyte differentiation to 3-dimensionally print electromechanically functional, chambered organoids composed of contiguous cardiac muscle. METHODS AND RESULTS: We optimized a photo-crosslinkable formulation of native ECM (extracellular matrix) proteins and used this bioink to 3-dimensionally print human induced pluripotent stem cell-laden structures with 2 chambers and a vessel inlet and outlet. After human induced pluripotent stem cells proliferated to a sufficient density, we differentiated the cells within the structure and demonstrated function of the resultant human chambered muscle pump. Human chambered muscle pumps demonstrated macroscale beating and continuous action potential propagation with responsiveness to drugs and pacing. The connected chambers allowed for perfusion and enabled replication of pressure/volume relationships fundamental to the study of heart function and remodeling with health and disease. CONCLUSIONS: This advance represents a critical step toward generating macroscale tissues, akin to aggregate-based organoids, but with the critical advantage of harboring geometric structures essential to the pump function of cardiac muscle. Looking forward, human chambered organoids of this type might also serve as a test bed for cardiac medical devices and eventually lead to therapeutic tissue grafting.
Authors: Karthikeyan Narayanan; Vivian Y Lim; Jiayi Shen; Zhen Wei Tan; Divya Rajendran; Shyh-Chyang Luo; Shujun Gao; Andrew C A Wan; Jackie Y Ying Journal: Tissue Eng Part A Date: 2013-10-17 Impact factor: 3.845
Authors: Harald C Ott; Thomas S Matthiesen; Saik-Kia Goh; Lauren D Black; Stefan M Kren; Theoden I Netoff; Doris A Taylor Journal: Nat Med Date: 2008-01-13 Impact factor: 53.440
Authors: Kavitha T Kuppusamy; Daniel C Jones; Henrik Sperber; Anup Madan; Karin A Fischer; Marita L Rodriguez; Lil Pabon; Wei-Zhong Zhu; Nathaniel L Tulloch; Xiulan Yang; Nathan J Sniadecki; Michael A Laflamme; Walter L Ruzzo; Charles E Murry; Hannele Ruohola-Baker Journal: Proc Natl Acad Sci U S A Date: 2015-05-11 Impact factor: 11.205
Authors: Yan Li; Archna Gautam; Jiwei Yang; Liqun Qiu; Zara Melkoumian; Jennifer Weber; Lavanya Telukuntla; Rashi Srivastava; Erik M Whiteley; Ralph Brandenberger Journal: Stem Cells Dev Date: 2013-02-13 Impact factor: 3.272
Authors: Beth Ripley; Tatiana Kelil; Michael K Cheezum; Alexandra Goncalves; Marcelo F Di Carli; Frank J Rybicki; Mike Steigner; Dimitrios Mitsouras; Ron Blankstein Journal: J Cardiovasc Comput Tomogr Date: 2015-12-12
Authors: Ilya Y Shadrin; Brian W Allen; Ying Qian; Christopher P Jackman; Aaron L Carlson; Mark E Juhas; Nenad Bursac Journal: Nat Commun Date: 2017-11-28 Impact factor: 14.919
Authors: Sangkyun Cho; Dennis E Discher; Kam W Leong; Gordana Vunjak-Novakovic; Joseph C Wu Journal: Nat Methods Date: 2022-09-05 Impact factor: 47.990
Authors: Jianhua Zhang; Zachery R Gregorich; Ran Tao; Gina C Kim; Pratik A Lalit; Juliana L Carvalho; Yogananda Markandeya; Deane F Mosher; Sean P Palecek; Timothy J Kamp Journal: Elife Date: 2022-06-27 Impact factor: 8.713
Authors: Marcus Alonso Cee Williams; Devin B Mair; Wonjae Lee; Esak Lee; Deok-Ho Kim Journal: Tissue Eng Part B Rev Date: 2021-03-16 Impact factor: 7.376