August 2018 - BNC-all - ecn.purdue.edu

Organ-on-a-Chip-Training-Workshop-September 13-14 2018
by Kwok, Tim 06 Aug '18

06 Aug '18

Dear Fellow Scientists, Recent advances in cell biology, microfabrication and microfluidics have enabled the development of microengineered and controlled models of the functional units of human organs — known as organs-on-a-chip or tissue-chips. The 3D3C workshop on organs-on-a-chip (initially scheduled in April, but postponed due to lecturer availability), will be offered on September 13 & 14, 2018. The workshop will include a lecture, classroom exercises, demonstrations and hands-on practical, covering the basic setup and modern approaches for the design of organs-on-a-chip. It will encompass aspects of substrates used in organs-on-a-chip, microfluidic systems, culture surface geometry and biosensors. During the lecture portion of the workshop we will also discuss differences between organoids and organs- or tissue-chips. For details, please refer to the attached pamphlet. The workshop is opened for enrollment and the deadline for registration is August 24, 2018. If you are interested, please contact me. Sincerely, Tim Kwok Facility Manager 3D Cell Culture Core (3D3C) Facility Birck Nanotechnology Center Purdue University ________________________________ [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/8967fb7a-c28…]

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Mailroom closed tomorrow until 10:30AM
by Totten, Mary J 06 Aug '18

06 Aug '18

Tomorrow Tuesday the 7th, the mailroom will be closed while the staff strip and wax the hallway. If you have packages to ship, please bring them today before 4:00PM. Otherwise your packages will not ship until Wednesday. Please let me know if you have any questions. Thank you, Mary Jo Totten Building Logisitcs Coordinator BirckNanotechnology Center/Purdue University 1205 West State Street West Lafayette, IN 47907-2057 (765) 496-1173

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3D3C Newsletter - June 2018
by Kwok, Tim 03 Aug '18

03 Aug '18

Dear All, Below please find the sixteenth issue of the newsletter of the 3D Cell Culture Core (3D3C) Facility of the Birck Nanotechnology Center. The newsletter is also available online (https://nanohub.org/groups/3d3cfacility/news) The four sections in the newsletter are: 3D at Purdue – this section highlights 3D cell culture-based research activity at Purdue 3D in focus – this section presents the current work on a specific 3D cell culture model or technique 3D in publications – this section brings a collection of recent publications on 3D cell culture 3D in meetings – this section includes a list of upcoming meetings related to 3D cell culture The newsletter will be available every two months. If you do not wish to receive the 3D3C newsletter in the future, please reply “cancel” to unsubscribe. Please contact me if you have questions. Yours Sincerely, Tim Kwok Facility Manager 3D Cell Culture Core (3D3C) Facility Birck Nanotechnology Center Purdue University ________________________________ [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/3e8e050f-67f…] Volume 16, June 2018 [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/10f15e72-d4d…] 3D at Purdue 3D in Focus 3D in Publications 3D in Meetings 3D at Purdue Blood-Brain Barrier Permeation Screening Since the pioneering work of Erlich (1985), the Blood-Brain Barrier (BBB) has fascinated investigators largely due to its ability to prevent many substrates from permeating into the brain parenchyma. After many years of research, it has been now established that brain capillaries are functionally and physiologically different from capillaries found in other parts of the central nervous system (CNS) and the body. A particularly important attribute of the BBB are the tight junctions lining the paracellular space against the brain microvessel endothelial cells (BMECs) of the capillary. They are comprised of distinct protein complexes that form a highly restrictive sieving barrier to the movement of most small and large hydrophilic molecules. Several significant differences exist between other CNS and systemic endothelia and the brain capillaries, including the fact that BMECs have minimal pinocytosis, possess a higher number of mitochondria (indicating higher metabolic requirements), express unique tight junctional proteins that restrict free paracellular diffusion. Moreover, BMECs are almost completely surrounded by astrocytic end feet and pericytes that provide structural support and work in synergy to form the neurovascular unit (Figure 1). In fact, most scientists support the idea that the complete neurovascular unit (NVU) is functionally the BBB, in contrast to the focus on BMECs alone. Therefore, BMECs in the NVU have a distinct environment that results in the formation of a physiologically dynamic barrier acting to restrict the free permeation of most compounds, including a majority of neurotherapeutic agents, from the blood into the neuronal parenchyma. Modeling permeation across the BBB to screen neurotherapeutic agents using simple and robust in vitro techniques has thus been a significant challenge to the field. [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/bf3ca789-c9e…] Figure 1. In vivo depiction of the BBB (left) in comparison to the in vitro BBB direct contact triculture with human neurons (right). Currently, most in vitro BBB screening methodologies involve seeding BMECs on a membranous TranswellTM filter support. In the monoculture configuration, the BMECs lack a physiologically relevant environment since it is without biochemical signaling or physical interaction with supporting NVU cells. In other cases, an effort to introduce the NVU relevant environment in in vitro models has been conducted by seeding either astrocytes or pericytes directly under the membranous filter support as a co-culture or as a triculture by additionally seeding astrocytes or pericytes on the bottom of the basal chamber. However, this approach does not allow optimal (and physiologically representative) direct interactions between astrocytes or pericytes and BMECs observed at the in vivo NVU level. While conventional indirect models have provided significant reduction in the permeation rates of hydrophilic paracellular markers, they still lack the extent of restriction and a physiologically representative BBB-configuration as found in vivo. We attribute this in part to the lack of direct contact between the BMECs, astrocytes and pericytes as found in the neurovascular unit, which we now realize is functionally the real blood-brain barrier. Recently, we have demonstrated in our laboratory (Kulczar et al., 2017) that we can directly layer BMECs on a confluent lawn of astrocytes seeded on the apical surface of a TranswellTM filter eliminating the barrier that exists in the indirect configuration. Through further research, we have developed a triculture model of the BBB that exhibits a more physiologically representative configuration of the BBB-system in vivo as illustrated in Figure 1. Contrary to all the published conventional methods, we have developed a triculture model of the BBB on the apical surface of a TranswellTM chamber in a way that allows all the principal cellular components of the BBB-system to interact as they do in their native in vivo environment, where BMECs are in direct contact with the astrocytes and at least partly with pericytes (Ngendahimana et al., 2017). Moreover, the basal well is available to contain cells of the brain parenchyma (e.g., neurons) or diseased cells (e.g., neuroblastoma cells) to investigate permeability-linked efficacy (ongoing research). Thus, we sought to develop this physiologically relevant configuration on the apical surface of a TranswellTM chamber mainly because: (1) Brain Microvessel Endothelial Capillary walls are not the only cells limiting drug entry into the brain parenchyma; the underlying astrocytes may also reduce drug permeability through both transcellular and paracellular routes, and (2) in vivo, pericytes form interdigitations with BMECs in order to modulate their expression of tight junctions and key enzymes. <https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/709df589-a87…> [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/bbb18c34-7a6…] <https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/709df589-a87…> <https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/709df589-a87…> Figure 2. Triculture of astrocytes, pericytes, and endothelial cells (hCMEC/D3). Through further research, we have developed a triculture model of the BBB that exhibits a more physiologically representative configuration of the BBB-system in vivo as illustrated in Figure 1. Contrary to all the published conventional methods, we have developed a triculture model of the BBB on the apical surface of a TranswellTM chamber in a way that allows all the principal cellular components of the BBB-system to interact as they do in their native in vivo environment, where BMECs are in direct contact with the astrocytes and at least partly with pericytes (Ngendahimana et al., 2017). Moreover, the basal well is available to contain cells of the brain parenchyma (e.g., neurons) or diseased cells (e.g., neuroblastoma cells) to investigate permeability-linked efficacy (ongoing research). Thus, we sought to develop this physiologically relevant configuration on the apical surface of a TranswellTM chamber mainly because: (1) Brain Microvessel Endothelial Capillary walls are not the only cells limiting drug entry into the brain parenchyma; the underlying astrocytes may also reduce drug permeability through both transcellular and paracellular routes, and (2) in vivo, pericytes form interdigitations with BMECs in order to modulate their expression of tight junctions and key enzymes. [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/6a071c41-0bf…]<https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/32f1a152-c57…> Figure 3. The rank ordering of established BBB permeants across the direct contact coculture model. Gregory Knipp, Ph.D. Associate Professor, Industrial and Physical Pharmacy Director of the Purdue Translational Pharmacology CTSI Core College of Pharmacy References Ehrlich P (1885) Das Sauerstoffbedürfnis des Organismus. In: Eine Farbenanalytische Studie, Hirschwald, Berlin. Kulczar C, Lubin KE, Lefebvre S, Miller DW, Knipp GT (2017) Development of a direct contact astrocyte-human cerebral microvessel endothelial cells blood-brain barrier coculture model. J Pharm Pharmacol, 69(12):1684-1696 Ngendahimana A, Kulczar CD, Lavan M, Lubin KE, Knipp GT. Patent Application No. 15/697,699. Entitled: “Blood Brain Barrier Models and Methods to Generate and Use the Same.” Submitted September 7, 2017, [https://patents.justia.com/patent/20180067103] ________________________________ 3D in Focus The composition and the organization of the extracellular matrix (ECM) are important regulators of cell behavior. These elements are also important for disease progression, like in cancer. Cells in proper 3D culture assemble in three-dimensional structures producing a physiologically relevant organization and function of tissues; however, this is only possible if the ECM is present. Recent studies on 3D culture have been focusing more on the behavior of cells than ECM composition. In cell culture the ECM is usually added exogenously as a start, but the understanding is that cells capable of matrix production will further establish their microenvironmental niche. Fibroblasts are among the essential producers of ECM in normal and diseased microenvironments. In the highlighted article, using mass spectrometry analysis, the ECM produced in cell culture of fibroblasts derived from prostate tissues and the in vivo matrisome from the same prostate stroma were compared. Ojalill M, Rappu P, Siljamäki E, Taimen P, Boström P, Heino J. The composition of prostate core matrisome in vivo and in vitro unveiled by mass spectrometric analysis. Prostate. 2018 Jun;78(8):583-594. doi: 10.1002/pros.23503. Epub 2018 Mar 9. 3D3C summary of the article: The cancerous and noncancerous lobes of prostate were surgically removed from six prostate cancer patients. Fibroblastic cell lines, one from the cancer tissue and one from the noncancerous lobe of each of the prostate tissues, were established in serum-free fibroblast basal medium. The ECM of cells cultured for seven days was extracted for proteome analysis by mass spectrometry. The matrisome of fibroblasts in culture derived from the stroma of cancerous and noncancerous lobes did not show differences in composition. When comparing the matrisome from the stroma of the noncancerous lobe of the corresponding prostate in vivo and the matrisome from cell culture, one-third of the proteins were found to be common. The majority of the proteins that were solely detected in samples in vivo were considered to be plasma derived. Most of the proteins solely found from in vitro fibroblast-derived ECM were related to matrix remodeling or growth factor action. To further study the effect of cell culture conditions on ECM production and composition, the matrisome from the monolayer culture of one fibroblast cell line was compared with the cells forced to form 3D aggregates (‘spheroids’ in micro-wells). ECM produced by fibroblasts in monolayer culture on plastic shared one-third of the proteins with the matrix produced by the same cells in spheroid culture, whereas more than one-third of the proteins were specific for each culture condition. In addition, the matrisome from spheroid culture did not resemble the in vivo ECM more closely than the matrisome from monolayer culture. Comments from 3D3C: The highlighted study attempts to address the question of whether fibroblasts in a 3D setting may recapitulate the ECM present in vivo by comparing ECM composition between prostate stromal tissue with fibroblasts derived from a similar tissue and cultured in monolayer and as spheroids. The approach to compare the matrisome from in vitro culture with corresponding in vivo prostate tissue is the major advantage of the study. However, it is important to note here that there is currently no appropriate 3D cell culture model for phenotypically normal prostate tissue, including the epithelium and the stroma. Producing spheroids is a standard and acceptable way to study tumors, but seeding only fibroblasts in micro-mold for spheroid culture does not mimic a physiologically relevant manner to reproduce the organization of these cells. Therefore, it might not be surprising that the composition of ECM produced by prostate derived fibroblasts in vitro from the ‘spheroid’ culture is not improved compared to that of the monolayer culture. These results support the necessity to improve 3D models for tissues that contain a stromal compartment, like the prostate. ________________________________ 3D in Publications Recent publications on 3D culture (please click to access the list on our web page https://nanohub.org/groups/3d3cfacility) Review<https://nanohub.org/groups/3d3cfacility/news#reviews> The research articles and reviews are arranged in the following categories: Scaffold free/Scaffold Organ/Tissue/Cell Others Spheroids<https://nanohub.org/groups/3d3cfacility/news#spheroids> Organoid<https://nanohub.org/groups/3d3cfacility/news#organoid> Scaffold<https://nanohub.org/groups/3d3cfacility/news#scaffold> Hydrogel<https://nanohub.org/groups/3d3cfacility/news#hydrogel> Matrix<https://nanohub.org/groups/3d3cfacility/news#matrix> Microfluidics<https://nanohub.org/groups/3d3cfacility/news#microfluidic> Microfabrication<https://nanohub.org/groups/3d3cfacility/news#microfabrication> Bone<https://nanohub.org/groups/3d3cfacility/news#bone> Bone Marrow<https://nanohub.org/groups/3d3cfacility/news#bonemarrow> Breast<https://nanohub.org/groups/3d3cfacility/news#breast> Colon<https://nanohub.org/groups/3d3cfacility/news#colon> Heart<https://nanohub.org/groups/3d3cfacility/news#heart> Liver<https://nanohub.org/groups/3d3cfacility/news#liver> Lung<https://nanohub.org/groups/3d3cfacility/news#lung> Muscle<https://nanohub.org/groups/3d3cfacility/news#muscle> Nerve<https://nanohub.org/groups/3d3cfacility/news#nerve> Prostate<https://nanohub.org/groups/3d3cfacility/news#prostate> Endothelial cells<https://nanohub.org/groups/3d3cfacility/news#endothelialcells> Fibroblast<https://nanohub.org/groups/3d3cfacility/news#fibroblast> Stem Cells<https://nanohub.org/groups/3d3cfacility/news#stemcells> Stromal Cells<https://nanohub.org/groups/3d3cfacility/news#stromalcells> Cancer/Tumor<https://nanohub.org/groups/3d3cfacility/news#cancer> Screening<https://nanohub.org/groups/3d3cfacility/news#screening> 3D bioprinting<https://nanohub.org/groups/3d3cfacility/news#3dbioprinting> Imaging<https://nanohub.org/groups/3d3cfacility/news#imaging> ________________________________ 3D in Meetings EMBL Conference: Microfluidics 2018: New Technologies and Applications in Biology, Biochemistry and Single-Cell Analysis Date: 15th to 17th July 2018 Location: Heidelberg, Germany Website: http://www.embl.de/training/events/2018/MCF18-01/index.html Contact person: events(a)embl.de<mailto:events@embl.de> Organized by: EMBL CCO Stem Cells & Tissue Engineering Summit 2018 Date: 18th to 20th July 2018 Location: Kaula Lumpur, Malaysia Website: https://www.stemcellssummit.org/stem-cells-summit Contact person: Karthik Telidevara Organized by: Subhadra Healthcare Gordon Research Conference — Signal Transduction by Engineered Extracellular Matrices Date: 22nd to 27th July 2018 Location: Andover, NH, USA Website: http://www.grc.org//signal-transduction-by-engineered-extracellular-matrice…<http://www.grc.org/signal-transduction-by-engineered-extracellular-matrices…> Contact person: Sarah C. Heilshorn Organized by: Gordon Research Conference PREDiCT: 3D Models Date: 21st to 23rd August 2018 Location: Boston, MA, USA Website: http://go.evvnt.com/224119-2?pid=80 Contact person: Doug Fairbrother Organized by: Hanson Wade 4th International Conference on Bio-based Polymers and Composites Date: 2nd to 6th September 2018 Location: Balatonfüred, Hungary Website: http://www.bipoco2018.hu Contact person: Dóra Tátraaljai Bio-based polymers and their blends, composites. Natural polymers and their modification. Natural fiber reinforced composites. Other raw materials based on natural resources. Biodegradation and environmental issues EMBO|EMBL Symposium: Organoids: Modelling Organ Development and Disease in 3D Culture Date: 10th to 13th September 2018 Location: Heidelberg, Germany Website: http://www.embo-embl-symposia.org/symposia/2018/EES18-08/index.html Contact person: events(a)embl.de<mailto:events@embl.de> Organized by: EMBL The 4Bio Summit Date: 27th to 28th November 2018 Location: Rotterdam, Netherlands Website: http://www.global-engage.com/event/4bio/?utm_source=4BIOEUMP2018 Contact person: Jane Williams Organized by: Global Engage Gordon Research Conference — Biomaterials and Tissue Engineering Date: 28th July to 2nd August 2019 Location: Castelldefels, Spain Website: http://www.grc.org//biomaterials-and-tissue-engineering-conference/2019/<http://www.grc.org/biomaterials-and-tissue-engineering-conference/2019/> Contact person: Jennifer L. West and Brendan A. Harley Organized by: Gordon Research Conference

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Closed Hallways Tuesday and Wednesday next week
by Totten, Mary J 02 Aug '18

02 Aug '18

Everyone, We will be closing hallways next week for stripping and waxing. Please make a note that you can still access the labs from the galley doors, but your offices will not be accessible until at least 11:00AM. Tuesday August 7th, BRK 1217 to 1239 hallway closed until 11:00AM Wednesday August 8th, BRK 1261 to 1273 hallway closed until 11:00AM If you have any questions, please contact me. Thank you, Mary Jo Totten Building Logisitcs Coordinator BirckNanotechnology Center/Purdue University 1205 West State Street West Lafayette, IN 47907-2057 (765) 496-1173

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Car Alarm - Red Dodge Journey
by Totten, Mary J 01 Aug '18

01 Aug '18

If you have a red Dodge Journey with a Massachusetts license 7LA 965, your car alarm has been going off for a couple of hours. Thank you, Mary Jo Totten Building Logisitcs Coordinator BirckNanotechnology Center/Purdue University 1205 West State Street West Lafayette, IN 47907-2057 (765) 496-1173

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