@article{d052afdc968b4aed8786d8735b90379f,
title = "Sulfonated cryogel scaffolds for focal delivery in ex-vivo brain tissue cultures",
abstract = "The human brain has unique features that are difficult to study in animal models, including the mechanisms underlying neurodevelopmental and psychiatric disorders. Despite recent advances in human primary brain tissue culture systems, the use of these models to elucidate cellular disease mechanisms remains limited. A major reason for this is the lack of tools available to precisely manipulate a specific area of the tissue in a reproducible manner. Here we report an easy-to-use tool for site-specific manipulation of human brain tissue in culture. We show that line-shaped cryogel scaffolds synthesized with precise microscale dimensions allow the targeted delivery of a reagent to a specific region of human brain tissue in culture. 3-sulfopropyl acrylate (SPA) was incorporated into the cryogel network to yield a negative surface charge for the reversible binding of molecular cargo. The fluorescent dyes BODIPY and DiI were used as model cargos to show that placement of dye loaded scaffolds onto brain tissue in culture resulted in controlled delivery without a burst release, and labelling of specific regions without tissue damage. We further show that cryogels can deliver tetrodotoxin to tissue, inhibiting neuronal function in a reversible manner. The robust nature and precise dimensions of the cryogel resulted in a user-friendly and reproducible tool to manipulate primary human tissue cultures. These easy-to-use cryogels offer an innovate approach for more complex manipulations of ex-vivo tissue.",
keywords = "Biomaterial tool, Human brain tissue, Local delivery, Mouse brain tissue, Neuronal function, Sulfonated cryogel, Tissue culture",
author = "Dimitri Eigel and Romy Schuster and M{\"a}nnel, {Max J.} and Julian Thiele and Panasiuk, {Martyna J.} and Andreae, {Laura C.} and Carmine Varricchio and Andrea Brancale and Welzel, {Petra B.} and Huttner, {Wieland B.} and Carsten Werner and Ben Newland and Long, {Katherine R.}",
note = "Funding Information: Human fetal brain tissue was obtained from the Human Development Biology Resource (HDBR), provided by the Joint MRC/Wellcome Trust (grant # 099175/Z/12/Z) Human Developmental Biology Resource (www.hdbr.org). The HDBR provided fresh tissue from fetuses aged 12?20 post-conception weeks (pcw): 12 pcw, n = 1; 14 pcw, n = 1; 17 pcw, n = 2; 20 pcw, n = 2. Human fetal brain tissue was dissected in PBS and used immediately for culture, cryogel experiments, or fixation. Cryogels were loaded with DiI, which was used at 2 mM, and/or CellTracker? Green BODIPY? Dye, which was used at 0.5 mM, then placed onto the tissue. Tissue was cultured as previously described [47]: placed into a rotating flask with 1.5 ml media outlined in 2.1, and incubated at 37 ?C in the presence of humidified 40% O2/5% CO2/55% N2. All tissue was fixed for at least 24 h at 4 ?C in 4% (wt/vol) paraformaldehyde (PFA) in 120 mM phosphate buffer (pH 7.4).The authors would like to thank the following funding sources: Wellcome Trust (Sir Henry Wellcome Postdoctoral Fellowship (BN)), the Deutsche Forschungsgemeinschaft (project number 320041273 (BN)) and the Medical Research Council [MR/S025065/1] (KL). MJP is supported by the UK Medical Research Council (grant number: 1934998) and King's College London is a member of the MRC Doctoral Training Partnership in Biomedical Sciences. JT receives funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (Grant agreement No. 852065). We would also like to thank Dianne Gerrelli, Steve Lisgo, Berta Crespo, and their teams at the HDBR for their invaluable support. Funding Information: The authors would like to thank the following funding sources: Wellcome Trust (Sir Henry Wellcome Postdoctoral Fellowship (BN)), the Deutsche Forschungsgemeinschaft (project number 320041273 (BN) ) and the Medical Research Council [ MR/S025065/1 ] (KL). MJP is supported by the UK Medical Research Council (grant number: 1934998 ) and King{\textquoteright}s College London is a member of the MRC Doctoral Training Partnership in Biomedical Sciences. JT receives funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (Grant agreement No. 852065 ). We would also like to thank Dianne Gerrelli, Steve Lisgo, Berta Crespo, and their teams at the HDBR for their invaluable support. Publisher Copyright: {\textcopyright} 2021 Elsevier Ltd Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",
year = "2021",
month = apr,
doi = "10.1016/j.biomaterials.2021.120712",
language = "English",
volume = "271",
journal = "Biomaterials",
issn = "0142-9612",
publisher = "Elsevier BV",
}
