TY - JOUR
T1 - Engineering anisotropic 3D tubular tissues with flexible thermoresponsive nanofabricated substrates
AU - Williams, Nisa P.
AU - Rhodehamel, Marcus
AU - Yan, Calysta
AU - Smith, Alec S.T.
AU - Jiao, Alex
AU - Murry, Charles E.
AU - Scatena, Marta
AU - Kim, Deok Ho
N1 - Funding Information:
We would like to thank the core facilities and staff of the Institute for Stem Cell and Regenerative Medicine (ISCRM), particularly the Tom and Sue Ellison Stem Cell Core and the Lynn and Mike Garvey Imaging Core. We thank Dr. Jonathan Tsui for extensive discussion and editing of this work. Dr. Eunpyo Choi was instrumental in the creation of graphics for this publication as well. This work was supported by the National Institutes of Health : R01HL146436 , UG3EB028094 , R01NS094388 , R01HL94388 (to D.H.K.), TR002317 (to A.S.T.S.), and 1F31HL145809-01A1 (to N.P.W.). The Human Frontier Science Program ( RGP0038/2018 to D.H.K). The Jaconette L. Tietze Young Scientist Award (to A.S.T.S.). Lastly, we are grateful for the institutional funding sources that supported this work: the Washington State funded ISCRM Fellows Program and the generous support from the Gree Real Estate company through the ISCRM Gree Scholars Program.
Funding Information:
We would like to thank the core facilities and staff of the Institute for Stem Cell and Regenerative Medicine (ISCRM), particularly the Tom and Sue Ellison Stem Cell Core and the Lynn and Mike Garvey Imaging Core. We thank Dr. Jonathan Tsui for extensive discussion and editing of this work. Dr. Eunpyo Choi was instrumental in the creation of graphics for this publication as well. This work was supported by the National Institutes of Health: R01HL146436, UG3EB028094, R01NS094388, R01HL94388 (to D.H.K.), TR002317 (to A.S.T.S.), and 1F31HL145809-01A1 (to N.P.W.). The Human Frontier Science Program (RGP0038/2018 to D.H.K). The Jaconette L. Tietze Young Scientist Award (to A.S.T.S.). Lastly, we are grateful for the institutional funding sources that supported this work: the Washington State funded ISCRM Fellows Program and the generous support from the Gree Real Estate company through the ISCRM Gree Scholars Program.
Publisher Copyright:
© 2020
PY - 2020/5
Y1 - 2020/5
N2 - Tissue engineering aims to capture the structural and functional aspects of diverse tissue types in vitro. However, most approaches are limited in their ability to produce complex 3D geometries that are essential for tissue function. Tissues, such as the vasculature or chambers of the heart, often possess curved surfaces and hollow lumens that are difficult to recapitulate given their anisotropic architecture. Cell-sheet engineering techniques using thermoresponsive substrates provide a means to stack individual layers of cells with spatial control to create dense, scaffold-free tissues. In this study, we developed a novel method to fabricate complex 3D structures by layering multiple sheets of aligned cells onto flexible scaffolds and casting them into hollow tubular geometries using custom molds and gelatin hydrogels. To enable the fabrication of 3D tissues, we adapted our previously developed thermoresponsive nanopatterned cell-sheet technology by applying it to flexible substrates that could be folded as a form of tissue origami. We demonstrated the versatile nature of this platform by casting aligned sheets of smooth and cardiac muscle cells circumferentially around the surfaces of gelatin hydrogel tubes with hollow lumens. Additionally, we patterned skeletal muscle in the same fashion to recapitulate the 3D curvature that is observed in the muscles of the trunk. The circumferential cell patterning in each case was maintained after one week in culture and even encouraged organized skeletal myotube formation. Additionally, with the application of electrical field stimulation, skeletal myotubes began to assemble functional sarcomeres that could contract. Cardiac tubes could spontaneously contract and be paced for up to one month. Our flexible cell-sheet engineering approach provides an adaptable method to recapitulate more complex 3D geometries with tissue specific customization through the addition of different cell types, mold shapes, and hydrogels. By enabling the fabrication of scaled biomimetic models of human tissues, this approach could potentially be used to investigate tissue structure-function relationships, development, and maturation in the dish.
AB - Tissue engineering aims to capture the structural and functional aspects of diverse tissue types in vitro. However, most approaches are limited in their ability to produce complex 3D geometries that are essential for tissue function. Tissues, such as the vasculature or chambers of the heart, often possess curved surfaces and hollow lumens that are difficult to recapitulate given their anisotropic architecture. Cell-sheet engineering techniques using thermoresponsive substrates provide a means to stack individual layers of cells with spatial control to create dense, scaffold-free tissues. In this study, we developed a novel method to fabricate complex 3D structures by layering multiple sheets of aligned cells onto flexible scaffolds and casting them into hollow tubular geometries using custom molds and gelatin hydrogels. To enable the fabrication of 3D tissues, we adapted our previously developed thermoresponsive nanopatterned cell-sheet technology by applying it to flexible substrates that could be folded as a form of tissue origami. We demonstrated the versatile nature of this platform by casting aligned sheets of smooth and cardiac muscle cells circumferentially around the surfaces of gelatin hydrogel tubes with hollow lumens. Additionally, we patterned skeletal muscle in the same fashion to recapitulate the 3D curvature that is observed in the muscles of the trunk. The circumferential cell patterning in each case was maintained after one week in culture and even encouraged organized skeletal myotube formation. Additionally, with the application of electrical field stimulation, skeletal myotubes began to assemble functional sarcomeres that could contract. Cardiac tubes could spontaneously contract and be paced for up to one month. Our flexible cell-sheet engineering approach provides an adaptable method to recapitulate more complex 3D geometries with tissue specific customization through the addition of different cell types, mold shapes, and hydrogels. By enabling the fabrication of scaled biomimetic models of human tissues, this approach could potentially be used to investigate tissue structure-function relationships, development, and maturation in the dish.
KW - 3D tissue engineering
KW - Cell-sheet engineering
KW - Nanofabrication
KW - Thermoresponsive polymer
UR - http://www.scopus.com/inward/record.url?scp=85079898577&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85079898577&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2020.119856
DO - 10.1016/j.biomaterials.2020.119856
M3 - Article
C2 - 32105818
AN - SCOPUS:85079898577
VL - 240
JO - Biomaterials
JF - Biomaterials
SN - 0142-9612
M1 - 119856
ER -