Generation of model tissues with dendritic vascular networks via sacrificial laser-sintered carbohydrate templates

ABSTRACT Sacrificial templates for patterning perfusable vascular networks in engineered tissues have been constrained in architectural complexity, owing to the limitations of

extrusion-based 3D printing techniques. Here, we show that cell-laden hydrogels can be patterned with algorithmically generated dendritic vessel networks and other complex hierarchical

networks by using sacrificial templates made from laser-sintered carbohydrate powders. We quantified and modulated gradients of cell proliferation and cell metabolism emerging in response to

fluid convection through these networks and to diffusion of oxygen and metabolites out of them. We also show scalable strategies for the fabrication, perfusion culture and volumetric

analysis of large tissue-like constructs with complex and heterogeneous internal vascular architectures. Perfusable dendritic networks in cell-laden hydrogels may help sustain thick and

densely cellularized engineered tissues, and assist interrogations of the interplay between mass transport and tissue function. Access through your institution Buy or subscribe This is a

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MULTISCALAR BIOLOGICAL FORMS Article 11 December 2024 LARGE-SCALE PERFUSED TISSUES VIA SYNTHETIC 3D SOFT MICROFLUIDICS Article Open access 12 January 2023 3D MICROMESH-BASED HYBRID

BIOPRINTING: MULTIDIMENSIONAL LIQUID PATTERNING FOR 3D MICROTISSUE ENGINEERING Article Open access 21 January 2022 DATA AVAILABILITY The main data supporting the results in this study are

available within the paper and its Supplementary Information. Much of the source and analysed data are available in Zenodo (https://doi.org/10.5281/zenodo.3723373). Some source datasets are

too large to be shared in public repositories and are available from the corresponding author on reasonable request. Design files and documentation for our open-source selective laser

sintering hardware and software are available in the Zenodo repository and at https://github.com/MillerLabFTW/OpenSLS. CODE AVAILABILITY A custom Python add-on for Blender to generate

bifurcating vascular structures is available in the Zenodo repository and at https://github.com/MillerLabFTW/IntussusceptionAddon. Image-processing and analysis scripts are also available in

the Zenodo repository. The mutual tree attraction algorithm for generating dendritic networks is closed source, but the generated architectures are included in the Zenodo repository. CHANGE

HISTORY * _ 16 JUNE 2021 A Correction to this paper has been published: https://doi.org/10.1038/s41551-021-00761-6 _ REFERENCES * Zamir, M. Fractal dimensions and multifractility in

vascular branching. _J. Theor. Biol._ 212, 183–190 (2001). CAS  PubMed  Google Scholar  * West, G. B., Brown, J. H. & Enquist, B. J. A general model for the origin of allometric scaling

laws in biology. _Science_ 276, 122–126 (1997). CAS  PubMed  Google Scholar  * West, G. B., Brown, J. H. & Enquist, B. J. A general model for ontogenetic growth. _Nature_ 413, 628–631

(2001). CAS  PubMed  Google Scholar  * Monahan-Earley, R., Dvorak, A. M. & Aird, W. C. Evolutionary origins of the blood vascular system and endothelium. _J. Thromb. Haemost._ 11, 46–66

(2013). PubMed  PubMed Central  Google Scholar  * Novosel, E. C., Kleinhans, C. & Kluger, P. J. Vascularization is the key challenge in tissue engineering. _Adv. Drug Deliv. Rev._ 63,

300–311 (2011). CAS  PubMed  Google Scholar  * Kinstlinger, I. S. & Miller, J. S. 3D-printed fluidic networks as vasculature for engineered tissue. _Lab Chip_ 16, 2025–2043 (2016). CAS 

PubMed  Google Scholar  * Cabodi, M. et al. A microfluidic biomaterial. _J. Am. Chem. Soc._ 127, 13788–13789 (2005). CAS  PubMed  Google Scholar  * Chrobak, K. M., Potter, D. R. & Tien,

J. Formation of perfused, functional microvascular tubes in vitro. _Microvasc. Res._ 71, 185–196 (2006). CAS  PubMed  Google Scholar  * Zhang, B. et al. Biodegradable scaffold with built-in

vasculature for organ-on-a-chip engineering and direct surgical anastomosis. _Nat. Mater._ 15, 669–678 (2016). CAS  PubMed  PubMed Central  Google Scholar  * Miller, J. S. The billion cell

construct: will three-dimensional printing get us there? _PLoS Biol._ 12, e1001882 (2014). PubMed  PubMed Central  Google Scholar  * Luo, Y., Lode, A. & Gelinsky, M. Direct plotting of

three-dimensional hollow fiber scaffolds based on concentrated alginate pastes for tissue engineering. _Adv. Healthc. Mater._ 2, 777–783 (2013). CAS  PubMed  Google Scholar  * Christensen,

K. et al. Freeform inkjet printing of cellular structures with bifurcations. _Biotechnol. Bioeng._ 112, 1047–1055 (2015). CAS  PubMed  Google Scholar  * Hinton, T. J. et al.

Three-dimensional printing of complex biological structures by freeform reversible embedding of suspended hydrogels. _Sci. Adv._ 1, e1500758 (2015). PubMed  PubMed Central  Google Scholar  *

Lee, A. et al. 3D bioprinting of collagen to rebuild components of the human heart. _Science_ 365, 482–487 (2019). CAS  PubMed  Google Scholar  * Zhang, R. & Larsen, N. B.

Stereolithographic hydrogel printing of 3D culture chips with biofunctionalized complex 3D perfusion networks. _Lab Chip_ 17, 4273–4282 (2017). CAS  PubMed  Google Scholar  * Meyer, W. et

al. Soft polymers for building up small and smallest blood supplying systems by stereolithography. _J. Funct. Biomater._ 3, 257–268 (2012). PubMed  PubMed Central  Google Scholar  *

Brandenberg, N. & Lutolf, M. P. In situ patterning of microfluidic networks in 3D cell-laden hydrogels. _Adv. Mater._ 28, 7450–7456 (2016). CAS  PubMed  Google Scholar  * Heintz, K. A.

et al. Fabrication of 3D biomimetic microfluidic networks in hydrogels. _Adv. Healthc. Mater._ 5, 2153–2160 (2016). CAS  PubMed  PubMed Central  Google Scholar  * Arakawa, C. K., Badeau, B.

A., Zheng, Y. & DeForest, C. A. Multicellular vascularized engineered tissues through user-programmable biomaterial photodegradation. _Adv. Mat._ 29, 1703156 (2017). Google Scholar  *

Grigoryan, B. et al. Functional intravascular topologies and multivascular networks within biocompatible hydrogels. _Science_ 364, 458–464 (2019). CAS  PubMed  PubMed Central  Google Scholar

  * Golden, A. P. & Tien, J. Fabrication of microfluidic hydrogels using molded gelatin as a sacrificial element. _Lab Chip_ 7, 720–725 (2007). CAS  PubMed  Google Scholar  * Miller, J.

S. et al. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. _Nat. Mater._ 11, 768–774 (2012). CAS  PubMed  PubMed Central  Google Scholar  *

Bégin-Drolet, A. et al. Design of a 3D printer head for additive manufacturing of sugar glass for tissue engineering applications. _Addit. Manuf._ 15, 29–39 (2017). Google Scholar  * Gelber,

M. K., Hurst, G., Comi, T. J. & Bhargava, R. Model-guided design and characterization of a high-precision 3D printing process for carbohydrate glass. _Addit. Manuf._ 22, 38–50 (2018).

CAS  Google Scholar  * Homan, K. A. et al. Bioprinting of 3D convoluted renal proximal tubules on perfusable chips. _Sci. Rep._ 6, 34845 (2016). CAS  PubMed  PubMed Central  Google Scholar 

* Kolesky, D. B. et al. 3D bioprinting of vascularized, heterogeneous cell-laden tissue constructs. _Adv. Mater._ 26, 3124–3130 (2014). CAS  PubMed  Google Scholar  * Kolesky, D. B., Homan,

K. A., Skylar-Scott, M. A. & Lewis, J. A. Three-dimensional bioprinting of thick vascularized tissues. _Proc. Natl Acad. Sci. USA_ 113, 3179–3184 (2016). CAS  PubMed  PubMed Central 

Google Scholar  * Skylar-Scott, M. A. et al. Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels. _Sci. Adv._ 5, eaaw2459 (2019). CAS  PubMed

  PubMed Central  Google Scholar  * Wu, W., Deconinck, A. & Lewis, J. A. Omnidirectional printing of 3D microvascular networks. _Adv. Mater._ 23, H178–H183 (2011). CAS  PubMed  Google

Scholar  * Song, K. H., Highley, C. B., Rouff, A. & Burdick, J. A. Complex 3D-printed microchannels within cell-degradable hydrogels. _Adv. Funct. Mater._ 28, 1801331 (2018). Google

Scholar  * Pimentel, C. R. et al. Three-dimensional fabrication of thick and densely populated soft constructs with complex and actively perfused channel network. _Acta Biomater._ 65,

174–184 (2018). Google Scholar  * Kinstlinger, I. S. et al. Open-source selective laser sintering (OpenSLS) of nylon and biocompatible polycaprolactone. _PLoS ONE_ 11, e0147399 (2016).

PubMed  PubMed Central  Google Scholar  * Roszelle, B. N. et al. Flow diverter effect on cerebral aneurysm hemodynamics: An in vitro comparison of telescoping stents and the Pipeline.

_Neuroradiology_ 55, 751–758 (2013). PubMed  Google Scholar  * Saggiomo, V. & Velders, A. H. Simple 3D printed scaffold-removal method for the fabrication of intricate microfluidic

devices. _Adv. Sci._ 2, 1500125 (2015). Google Scholar  * Nguyen, L. H. et al. Vascularized bone tissue engineering: approaches for potential improvement. _Tissue Eng. Part B_ 18, 363–382

(2012). CAS  Google Scholar  * Nguyen, Q. T., Hwang, Y., Chen, A. C., Varghese, S. & Sah, R. L. Cartilage-like mechanical properties of poly (ethylene glycol)-diacrylate hydrogels.

_Biomaterials_ 33, 6682–6690 (2012). CAS  PubMed  PubMed Central  Google Scholar  * Partlow, B. P. et al. Highly tunable elastomeric silk biomaterials. _Adv. Funct. Mater._ 24, 4615–4624

(2014). CAS  PubMed  PubMed Central  Google Scholar  * Mooney, R., Tawil, B. & Mahoney, M. Specific fibrinogen and thrombin concentrations promote neuronal rather than glial growth when

primary neural cells are seeded within plasma-derived fibrin gels. _Tissue Eng. Part A_ 16, 1607–1619 (2010). CAS  PubMed  Google Scholar  * Duong, H., Wu, B. & Tawil, B. Modulation of

3D fibrin matrix stiffness by intrinsic fibrinogen–thrombin compositions and by extrinsic cellular activity. _Tissue Eng. Part A_ 15, 1865–1876 (2009). CAS  PubMed  PubMed Central  Google

Scholar  * Subhash, G., Liu, Q., Moore, D. F., Ifju, P. G. & Haile, M. A. Concentration dependence of tensile behavior in agarose gel using digital image correlation. _Exp. Mech._ 51,

255–262 (2011). CAS  Google Scholar  * Feugier, F. G., Mochizuki, A. & Iwasa, Y. Self-organization of the vascular system in plant leaves: inter-dependent dynamics of auxin flux and

carrier proteins. _J. Theor. Biol._ 236, 366–375 (2005). CAS  PubMed  Google Scholar  * Fujita, H. & Mochizuki, A. The origin of the diversity of leaf venation pattern. _Dev. Dyn._ 235,

2710–2721 (2006). PubMed  Google Scholar  * Runions, A., Lane, B. & Prusinkiewicz, P. Modeling trees with a space colonization algorithm. In _Proc. 3rd Eurographics Conference on Natural

Phenomena_ (Eds Ebert, D. & Mérillou, S.) 63–70 (Eurographics Association, 2007). * Murray, C. D. The physiological principle of minimum work applied to the angle of branching of

arteries. _J. Gen. Physiol._ 9, 835–841 (1926). CAS  PubMed  PubMed Central  Google Scholar  * Miguel, A. F. Dendritic design as an archetype for growth patterns in nature: fractal and

constructal views. _Front. Phys._ 2, 9 (2014). Google Scholar  * Moon, J. J. et al. Biomimetic hydrogels with pro-angiogenic properties. _Biomaterials_ 31, 3840–3847 (2010). CAS  PubMed 

PubMed Central  Google Scholar  * Calderon, G. et al. Tubulogenesis of co-cultured human iPS-derived endothelial cells and human mesenchymal stem cells in fibrin and gelatin methacrylate

gels. _Biomater. Sci._ 5, 1652–1660 (2017). CAS  PubMed  Google Scholar  * Eskin, S. G., Ives, C., McIntire, L. & Navarro, L. Response of cultured endothelial cells to steady flow.

_Microvasc. Res._ 28, 87–94 (1984). CAS  PubMed  Google Scholar  * Yang, P. J. & Temenoff, J. S. Engineering orthopedic tissue interfaces. _Tissue Eng. Part B_ 15, 127–141 (2009). CAS 

Google Scholar  * Eckes, B. et al. Fibroblast-matrix interactions in wound healing and fibrosis. _Matrix Biol._ 19, 325–332 (2000). CAS  PubMed  Google Scholar  * Lu, P., Weaver, V. M. &

Werb, Z. The extracellular matrix: a dynamic niche in cancer progression. _J. Cell Biol._ 196, 395–406 (2012). CAS  PubMed  PubMed Central  Google Scholar  * Radisic, M. et al. Oxygen

gradients correlate with cell density and cell viability in engineered cardiac tissue. _Biotechnol. Bioeng._ 93, 332–343 (2006). CAS  PubMed  Google Scholar  * Tocchio, A. et al. Versatile

fabrication of vascularizable scaffolds for large tissue engineering in bioreactor. _Biomaterials_ 45, 124–131 (2015). CAS  PubMed  Google Scholar  * Tsang, V. L. et al. Fabrication of 3D

hepatic tissues by additive photopatterning of cellular hydrogels. _FASEB J._ 21, 790–801 (2007). CAS  Google Scholar  * Krogh, A. The number and distribution of capillaries in muscles with

calculations of the oxygen pressure head necessary for supplying the tissue. _J. Physiol._ 52, 409–415 (1919). CAS  PubMed  PubMed Central  Google Scholar  * Lewis, M. C., MacArthur, B. D.,

Malda, J., Pettet, G. & Please, C. P. Heterogeneous proliferation within engineered cartilaginous tissue: the role of oxygen tension. _Biotechnol. Bioeng._ 91, 607–615 (2005). CAS 

PubMed  Google Scholar  * Demol, J., Lambrechts, D., Geris, L., Schrooten, J. & Van Oosterwyck, H. Towards a quantitative understanding of oxygen tension and cell density evolution in

fibrin hydrogels. _Biomaterials_ 32, 107–118 (2011). CAS  PubMed  Google Scholar  * Gu, W. Y., Yao, H., Huang, C. Y. & Cheung, H. S. New insight into deformation-dependent hydraulic

permeability of gels and cartilage, and dynamic behavior of agarose gels in confined compression. _J. Biomech._ 36, 593–598 (2003). CAS  PubMed  Google Scholar  * Chuppa, S. et al. Fermentor

temperature as a tool for control of high-density perfusion cultures of mammalian cells. _Biotechnol. Bioeng._ 55, 328–338 (1997). CAS  PubMed  Google Scholar  * Ducommun, P., Ruffieux, P.

A., Kadouri, A., Von Stockar, U. & Marison, I. W. Monitoring of temperature effects on animal cell metabolism in a packed bed process. _Biotechnol. Bioeng._ 77, 838–842 (2002). CAS 

PubMed  Google Scholar  * Jorjani, P. & Ozturk, S. S. Effects of cell density and temperature on oxygen consumption rate for different mammalian cell lines. _Biotechnol. Bioeng._ 64,

349–356 (1999). CAS  PubMed  Google Scholar  * Xiang, C. et al. Long-term functional maintenance of primary human hepatocytes in vitro. _Science_ 364, 399–402 (2019). CAS  PubMed  Google

Scholar  * Bhatia, S. N., Balis, U. J., Yarmush, M. L. & Toner, M. Effect of cell–cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal

cells. _FASEB J._ 13, 1883–1900 (1999). CAS  PubMed  Google Scholar  * Stevens, K. R. et al. InVERT molding for scalable control of tissue microarchitecture. _Nat. Commun._ 4, 1847 (2013).

CAS  PubMed  Google Scholar  * Stevens, K. R. et al. In situ expansion of engineered human liver tissue in a mouse model of chronic liver disease. _Sci. Transl. Med._ 9, eaah5505 (2017).

PubMed  PubMed Central  Google Scholar  * Khetani, S. R. & Bhatia, S. N. Microscale culture of human liver cells for drug development. _Nat. Biotechnol._ 26, 120–126 (2008). CAS  PubMed

  Google Scholar  * Bhatia, S. N., Underhill, G. H., Zaret, K. S. & Fox, I. J. Cell and tissue engineering for liver disease. _Sci. Transl. Med._ 6, 245sr2 (2014). PubMed  PubMed Central

  Google Scholar  * Rafii, S., Butler, J. M. & Ding, B.-S. Angiocrine functions of organ-specific endothelial cells. _Nature_ 529, 316–325 (2016). CAS  PubMed  PubMed Central  Google

Scholar  * Baranski, J. D. et al. Geometric control of vascular networks to enhance engineered tissue integration and function. _Proc. Natl Acad. Sci USA_ 110, 7586–7591 (2013). CAS  PubMed

  PubMed Central  Google Scholar  * Mirabella, T. et al. 3D-printed vascular networks direct therapeutic angiogenesis in ischaemia. _Nat. Biomed. Eng._ 1, 0083 (2017). CAS  PubMed  PubMed

Central  Google Scholar  * Lindström, N. O. et al. Conserved and divergent molecular and anatomic features of human and mouse nephron patterning. _J. Am. Soc. Nephrol._ 29, 825–840 (2018).

PubMed  PubMed Central  Google Scholar  * Bosco, D. et al. Unique arrangement of α- and β-cells in human islets of Langerhans. 59, 1202–1210 (2010). * Wang, X.-N. et al. A three-dimensional

atlas of human dermal leukocytes, lymphatics, and blood vessels. _J. Invest. Dermatol._ 134, 965–974 (2014). CAS  PubMed  Google Scholar  * Kang, H.-W. et al. A 3D bioprinting system to

produce human-scale tissue constructs with structural integrity. _Nat. Biotechnol._ 34, 312–319 (2016). CAS  PubMed  Google Scholar  * Shapiro, A. J. et al. Islet transplantation in seven

patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regime. _N. Engl. J. Med._ 343, 230–238 (2000). CAS  PubMed  Google Scholar  * Iansante, V., Mitry, R.

R., Filippi, C., Fitzpatrick, E. & Dhawan, A. Human hepatocyte transplantation for liver disease: current status and future perspectives. _Pediatr. Res._ 83, 232–240 (2018). CAS  PubMed

  Google Scholar  * Parker Ponder, K. et al. Mouse hepatocytes migrate to liver parenchyma and function indefinitely after intrasplenic transplantation. _Proc. Natl Acad. Sci. USA_ 88,

1217–1221 (1991). Google Scholar  * Truslow, J. G. & Tien, J. Perfusion systems that minimize vascular volume fraction in engineered tissues. _Biomicrofluidics_ 5, 022201 (2011). PubMed

Central  Google Scholar  * Ronellenfitsch, H. & Katifori, E. Global optimization, local adaptation, and the role of growth in distribution networks. _Phys. Rev. Lett._ 117, 138301

(2016). PubMed  Google Scholar  * Freeman, R. Measuring the flow properties of consolidated, conditioned and aerated powders—a comparative study using a powder rheometer and a rotational

shear cell. _Powder Technol._ 174, 25–33 (2007). CAS  Google Scholar  * Thadavirul, N., Pavasant, P. & Supaphol, P. Development of polycaprolactone porous scaffolds by combining solvent

casting, particulate leaching, and polymer leaching techniques for bone tissue engineering. _J. Biomed. Mater. Res. A_ 102, 3379–3392 (2013). PubMed  Google Scholar  * Miller, J. S. et al.

Bioactive hydrogels made from step-growth derived PEG-peptide macromers. _Biomaterials_ 31, 3736–3743 (2010). CAS  PubMed  PubMed Central  Google Scholar  * Rockwood, D. N. et al. Materials

fabrication from _Bombyx mori_ silk fibroin. _Nat. Protoc._ 6, 1612–1631 (2011). CAS  PubMed  Google Scholar  * Li, W., Germain, R. N. & Gerner, M. Y. Multiplex, quantitative cellular

analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D)._Proc. Natl Acad. Sci. USA_ 114, E7321–E7330 (2017). CAS  PubMed  PubMed Central  Google Scholar  * Thielicke, W.

& Stamhuis, E. J. PIVlab—towards user-friendly, affordable and accurate digital particle image velocimetry in MATLAB. _J. Open Res. Softw._ 2, e30 (2014). Google Scholar  * Thielicke,

W. _The Flapping Flight of Birds—Analysis and Application_. PhD thesis, Rijksuniversiteit Groningen (2014). * Thielicke, W. & Stamhuis, E. J. PIVlab—time-resolved digital particle image

velocimetry tool for MATLAB https://doi.org/10.6084/M9.FIGSHARE.1092508.V5 (2014). * Cheng, N.-S. Formula for the viscosity of a glycerol–water mixture. _Ind. Eng. Chem. Res._ 47, 3285–3288

(2008). CAS  Google Scholar  * Volk, A. & Kähler, C. J. Density model for aqueous glycerol solutions. _Exp. Fluids_ 59, 75 (2018). Google Scholar  * van de Loosdrecht, A. A., Beelen, R.

H., Ossenkoppele, G. J., Broekhoven, M. G. & Langenhuijsen, M. M. A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells

from cell lines and patients with acute myeloid leukemia. _J. Immunol. Methods_ 174, 311–320 (1994). PubMed  Google Scholar  * Ahrens, J., Geveci, B. & Law, C. in _The Visualization

Handbook_ (eds Hansen, C. D. & Johnson, C. R.) 717–731 (2005). Download references ACKNOWLEDGEMENTS We thank A. Bastian, A. Ta and T. Schmidt for assistance with OpenSLS hardware and

firmware; E. Watson and A. Mikos for assistance with mechanical testing; J. Wagner, P. Desai and C. F. Higgs for assistance with powder rheology; D. De Santos for technical assistance with

carbohydrate SLS; D. Kaplan and W. Stoppel for providing silk fibroin; D. L. Gibbons for providing the 344SQ lung adenocarcinoma cell line; and C. Fortin for help with hepatocyte isolations.

This work was supported in part by a Medical Research Grant from the Robert J. Kleberg Jr and Helen C. Kleberg Foundation (J.S.M.), National Institutes of Health (grants HL134510 and

DK115461 (K.-D.B.)), the Texas Hepatocellular Carcinoma Consortium (THCCC) (CPRIT RP150587 (K.-D.B.)), National Insitutes of Health grant DP2HL137188 (K.R.S.) and National Insitutes of

Health NIBIB Cardiovascular Training grant (T32EB001650 (S.H.S.)). I.S.K. acknowledges support by an F31 National Research Service Award (NRSA) from the National Institutes of Health

(HL140905). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS *

Department of Bioengineering, Rice University, Houston, TX, USA Ian S. Kinstlinger, Gisele A. Calderon, Karen Vasquez Ruiz, David R. Yalacki, Palvasha R. Deme, Kevin D. Janson, Daniel W.

Sazer, Saarang S. Panchavati & Jordan S. Miller * Department of Bioengineering, University of Washington, Seattle, WA, USA Sarah H. Saxton, Fredrik Johansson & Kelly R. Stevens *

Nervous System, Palenville, NY, USA Jessica E. Rosenkrantz & Jesse D. Louis-Rosenberg * Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA

Karl-Dimiter Bissig * Department of Pathology, University of Washington, Seattle, WA, USA Kelly R. Stevens Authors * Ian S. Kinstlinger View author publications You can also search for this

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Scholar CONTRIBUTIONS I.S.K. and J.S.M. conceived and initiated the project and wrote the manuscript. I.S.K., S.H.S., G.A.C., K.V.R., D.R.Y., P.R.D., K.D.J. and F.J. designed and performed

experiments. I.S.K., S.H.S. and G.A.C. acquired and analysed imaging data. J.E.R., J.D.L.-R. and S.S.P. developed generative design algorithms. D.W.S. synthesized materials. K.-D.B., K.R.S.

and J.S.M. supervised the project. CORRESPONDING AUTHOR Correspondence to Jordan S. Miller. ETHICS DECLARATIONS COMPETING INTERESTS J.S.M. is a co-founder and holds an equity stake in

Volumetric, Inc. J.E.R. and J.D.L.-R. are co-founders and hold equity stakes in Nervous System, Inc. ADDITIONAL INFORMATION PUBLISHER’S NOTE Springer Nature remains neutral with regard to

jurisdictional claims in published maps and institutional affiliations. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary Methods. REPORTING SUMMARY SUPPLEMENTARY VIDEO 1

Annotated video recording of one layer of carbohydrate SLS. SUPPLEMENTARY VIDEO 2 Mutual-tree-attraction algorithm for the generative design of dendritic networks. SUPPLEMENTARY VIDEO 3

Fluorescent bead perfusion through a whole planar dendritic network. SUPPLEMENTARY VIDEO 4 Magnified view of fluorescent bead perfusion through the centre of a planar dendritic network.

SUPPLEMENTARY VIDEO 5 Perfusion of dendritic architectures at a high volumetric flow rate. SUPPLEMENTARY VIDEO 6 Animated volume rendering of a region in an endothelialized planar dendritic

network. SUPPLEMENTARY VIDEO 7 Rotating rendering of a volumetric μCT scan of a dendritic carbohydrate template. SUPPLEMENTARY VIDEO 8 Fly-through rendering of a volumetric μCT scan of a

dendritic carbohydrate template. SUPPLEMENTARY VIDEO 9 Fly-through sequence of MTT staining in sections from a cell-laden gel with dendritic architecture. SUPPLEMENTARY VIDEO 10 Fly-through

sequence of nuclear staining in sections from a cell-laden gel with dendritic architecture. SUPPLEMENTARY VIDEO 11 Fly-through sequence of processed MTT staining images from a cell-laden gel

with dendritic architecture. SUPPLEMENTARY VIDEO 12 Rotating rendering of a volumetrically reconstructed MTT signal in a cell-laden gel with dendritic architecture. SUPPLEMENTARY VIDEO 13

Computational fluid-dynamics simulation of perfusion through a 3D dendritic architecture. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Kinstlinger,

I.S., Saxton, S.H., Calderon, G.A. _et al._ Generation of model tissues with dendritic vascular networks via sacrificial laser-sintered carbohydrate templates. _Nat Biomed Eng_ 4, 916–932

(2020). https://doi.org/10.1038/s41551-020-0566-1 Download citation * Received: 27 September 2018 * Accepted: 01 May 2020 * Published: 29 June 2020 * Issue Date: September 2020 * DOI:

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