You can select the "Continue Account Application" button below if you need to complete your application. Studies of brain cell function and physiology are hampered by the limited availability of immortal human brain-derived cell lines, as a result of the technical difficulties encountered in establishing immortal human cells in culture. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. * Keep the medium to surface area ratio at 1mL per 5 cm 2. The cells were digested with EDTA-free trypsin, centrifuged at 1000g for 5min, then re-suspended with 1ml PBS and counted. For more information, please browse the options below to take a deeper look at the tools for your research needs. Whittemore SR, Neary JT, Kleitman N, Sanon HR, Benigno A, Donahue RP, Norenberg MD. WebHuman Astrocytes Reference: P10251 Human Astrocytes (HA) provided by Innoprot are isolated from human healthy brain (cerebral cortex). 5C). To download a certificate of analysis for C8-D1A [Astrocyte type I clone] (CRL-2541), enter the lot number exactly as it appears on your product label or packing slip. As shown in Fig. physical and metabolic support for the normal functioning of neurons and other neuronal cells. The result showed that the highest exosome diameter was about 98nm. 4 D) (Oberheim All Rights Reserved. They could also stimulate astrocytes to secrete numerous cytokines that promote the formation of the inflammatory and immune inhibitory microenvironment. A Relative molecular weight distribution of differentially expressed proteins in SVG P12 cells. With our integrated global resources, our services and products will provide you with a cutting-edge platform that could exponentially speed up the process of neurology research, and ultimately promote science for the cause of a better tomorrow. Drug Discov Today Dis Models. HIV-1, hepatitis B and hepatitis C are not detected for all donors and/or cell lots. HA are cryopreserved at passage one and delivered frozen. Astrocytes facilitate melanoma brain metastasis via secretion of IL-23. How Does Cre, Flp, Dre Recombinase System Work? WebPrimary human astrocytes were transfected with the SV40 T antigen expression vectors, pSV3neo or p735.6, and cultures were established with an extended lifespan. -. Up to now, the central nervous system has been considered as the refuge of brain metastases. Nat Cell Biol. 5D). Normal Human Astrocytes (NHA) can be used to study the function of the central nervous system and how neural cells interact. They are involved in various cognitive functions like sleep and memory formation. Studies of brain cell function and physiology are hampered by the limited availability of immortal human brain-derived cell lines, as a result of the technical difficulties encountered in establishing immortal human cells in culture. Ferroptosis: a potential opportunity for intervention of pre-metastatic niche. J Pathol. volume25, Articlenumber:5 (2023) The vertical axis showed the name of each GO function, and the value in horizontal axis was the number of differentially expressed proteins contained in that function. 2021 May;200:101973. doi: 10.1016/j.pneurobio.2020.101973. Creative Biolabs's product portfolio also includes reliable cell differentiation media, optimized serum-free culture media, medium additives, key cell markers and research antibodies. MeSH Epub 2018 Aug 9. WebThe ATCC collection has many neural cell lines, representing the normal and diseased tissue of multiple species such as neurons and the supporting cells of the central and Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. Furthermore, this cell line was cultivated in presence of human astrocyte medium as the barrier properties are mostly induced and maintained by the close links between brain endothelial cells and astrocytes (Abbott et al., 2006, Janzer and Raff, 1987). Accessibility GRO/CXCL1, IFN-, IL-3, IL-5, IL-15, LIF, M-CSF, NGF, PDGF, and VEGF secreted by SVG P12 cells were significantly up-regulated, while IL-7 secretion was significantly reduced. 2019;18(1):39. To summarize, HASTR/ci35 cells possess several key astrocytic characteristics, including various transporter functions, while simultaneously showing infinite proliferation and scalability. Semin Cancer Biol. Wang M, Qin Z, Wan J, et al. When co-cultured with neuronal cells, HASTR/ci35 cells could facilitate their dendritic network formation. For both iPast lines, the expression level of SLC1A3 in A4-step had decreased to the level in F A. PubMed Type I and II cells had small somata. Nat Rev Cancer. Disclaimer. 8:e80970. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Lingyun Ye wrote the manuscript and carried out some experiments. 2021;21(1):556. Astrocytes are the most plentiful cells in the human brain and play We have received your request for this certificate of origin. All of the operations from this point on should be carried out under strict aseptic conditions. However, the effect of lung cancer cell-derived exosomes on astrocytes during the formation of brain PMN in lung cancer brain metastasis had not been reported so far. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. The collection includes normal Once UTSW63 and UTSW71 cell lines were established, only low passage cells were used. WebHuman primary cells more closely mimic the physiological state of cells in vivo and generate more relevant data representing living systems. government site. Carcinogenesis. PubMed Unable to load your collection due to an error, Unable to load your delegates due to an error. Primary human astrocytes were transfected with the SV40 T antigen expression vectors, pSV3neo or p735.6, and cultures were established with an extended lifespan. After incubation for 24h, the Edu working solution was prepared according to the instructions. 1321N1 is a human astrocytoma cell line isolated in 1972 as a sub clone of the cell line 1181N1 which in turn was isolated from the parent line U-118 MG (one of a number of cell lines derived from malignant gliomas by J Ponten .1321N1 has been shown to have very similar STR profile data to U-118 MG. In fact, an interesting RNAseq study (Zhang et al., 2016) revealed that mature human astrocytes derived from the temporal lobe cortex are the primary cell type expressing After Assay Buffer was used to dilute and shake microbeads, 50l was taken and added to a 96-well plate, followed by 50l standard, sample, and Blank, and the samples were incubated at room temperature for 30min without light. The product is provided 'AS IS' and the viability of ATCC products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. Fluids Barriers CNS. Most significantly, human astrocytes extended 10-fold more GFAP + processes from their cell body than do rodent astrocytes, with an average of 37.5 5.17 processes radiating from the nucleus compared with 3.75 0.115 in the rodent (human, n = 50 cells from 7 patients; rodent, n = 65 cells 7 animals; p < 0.0001) (Fig. P3 Primary Cell 96-well Nucleofector Kit, P3 Primary Cell 96-well Nucleofector Kit (960 RCT), P3 Primary Cell 384-well Nucleofector Kit, Liver Cells for ADME & Toxicology Studies, Endotoxin Detection and Analysis Software, MODA-ES The Next Generation Electronic Batch, MES for Cell and Gene Therapy Manufacturing, Guaranteed through 10 population doublings, Positive immunofluorescence staining for glial fibrillary acid protein (GFAP), Test negative for mycoplasma, bacteria, yeast, and fungi, HIV-1, Hepatitis B and Hepatitis C are not detected for all donors and/or cell lots, A Certificate ofAnalysis is provided for each cell lot purchased. In addition, the proteomic analysis showed that H1299 cell-derived exosomes changed the expression of the proteins which were mainly involved in regulating metabolic pathways in astrocytes. Morrissey SM, Zhang F, Ding C, et al. 2015;527(7578):329. WebProduct Overview. 7). Astrocyte development: A Guide for the Perplexed. 2007 Jan;210(1):54-65. doi: 10.1111/j.1469-7580.2006.00671.x. CD9, CD63, HSP90, and Alix were used as exosome-positive protein markers, and Calnexin as an exosome-negative protein marker. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. E Statistical analysis of flow cytometry results (p<0.0001). Tumor-derived exosomes drive immunosuppressive macrophages in a pre-metastatic niche through glycolytic dominant metabolic reprogramming. Clin Cancer Res. Exo: SVG P12 cells co-cultured with H1299 cell-derived exosomes for 24h). It seems that loss of astrocytes will destroy BBB and promote the formation of PMN, therefore further studies are required to clarify the underlying mechanisms. https://doi.org/10.1186/s12575-023-00192-4, DOI: https://doi.org/10.1186/s12575-023-00192-4. We conclude that methods based on the use of SV40 T antigen can successfully immortalize human astrocytes, retaining key astrocyte functions, but T antigen-induced proliferation appeared to interfere with expression of glial fibrillary acidic protein. Tan HY, Yong YK, Xue YC, Liu H, Furihata T, Shankar EM, Ng CS. We believe A735 is the first documented nontumor-derived human glial cell line which is immortal. Would you like email updates of new search results? 2014;156:100216. Your application may be in one of the following statuses: You can check the status of your application in your My Dashboard portal. Cecchelli R, Berezowski V, Lundquist S, et al. The authors declare no competing interests. Meanwhile, apoptosis-related proteins MAP2K1, TUBA1C, RELA, and CASP6 were up-regulated. WebFigure Legend Snippet: Tha-eGFP and Th2P-4M-eGFP preferentially replicate and spread in human SK-N-SH compared to astrocyte-like (SVGp12) and microglia-like (HMC3) cell lines in vitro. J. We present a three-dimensional induced pluripotent stem cell (iPSC)-derived paradigmmature brain organoids (mbOrg)composed of cortical-like-astrocytes (iA) and neurons. The 96-well plates were placed in an incubator for 24hours and removed. Animal in March 1993. After chromatographic separation by the Easy nLC system, the samples were analyzed with Orbitrap Exploris 480 mass spectrometer. At last, the Bio-Plex detector was used to detect the concentration of each sample. We will not share your information outside of our distributors network and solely use it to send relevant communications. Third row, commercially available cell lines grown in serum-containing media, from patients with verified disease states. Tumor cells could also activate cGAMP in STAT1 and NF-B signaling pathways to form gap junctions with astrocytes and stimulate the latter to produce IFN- and other cytokines, which in turn support tumor cell growth [22]. Therefore, our results suggest that the age-associated upregulation of GS and GLT-1 is a conserved feature between mouse and human hippocampal astrocytes. 2021 Apr 16;22(8):4116. doi: 10.3390/ijms22084116. ( A ) Schematic overview of the genomic organization of Tha-eGFP and Th2P-4M-eGFP. Overall, our study provides a new theoretical basis for lung cancer brain metastasis. The exosomes were diluted to 10^7 / ml-10 ^9 /ml with PBS, and the size and quality of the exosomes were measured by NanoSight NS300 particle potentiometric titration and particle size analyzer. PubMed Central Biological Procedures Online It is your responsibility to understand the hazards associated with the material per your organizations policies and procedures as well as any other applicable regulations as enforced by your local or national agencies. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. Bethesda, MD 20894, Web Policies Fong MY, Zhou W, Liu L, et al. Das M, Smith ML, Furihata T, Sarker S, O'Shea R, Helbig KJ. 2021;2(1):3. Alliot F, Pessac B. Astrocytic cell clones derived from established cultures of 8-day postnatal mouse cerebella. Parenchymal astrocytes retain a latent neurogenic program, which can be reactivated by brain injury. 2010 Nov;6(4):263-76. doi: 10.1017/S1740925X11000093. Correspondence to Modelling of the blood-brain barrier in drug discovery and development. 2016;30(5):66881. PubMedGoogle Scholar. In our study, we used the label-free technique to determine the effect of exosomes secreted by H1299 cells on the protein expression of astrocytes. Tha-eGFP and Th2P-4M-eGFP harbour the same genetic background except Generation of two gene edited iPSC-lines carrying a DOX-inducible NGN2 expression cassette with and without GFP in the AAVS1 locus. are derived from human cerebral cortex. 2015;6:6716. FOIA Indeed, our 3D iPSC-induced cortical-like neuron and astrocyte co-culture with GRN loss strikingly and consistently recapitulated human specific TDP-43 cell pathology reported in human FTD patient brain. Orr FW, Wang HH, Lafrenie RM, et al. Piantino M, Kang DH, Furihata T, Nakatani N, Kitamura K, Shigemoto-Mogami Y, Sato K, Matsusaki M. Mater Today Bio. And when the concentration of exosomes reached 60g/ml, the viability of SVG P12 cells decreased by about 50%. 2020 Jul 16;17(1):44. doi: 10.1186/s12987-020-00202-7. And the differentially expressed proteins were involved in regulating metabolic pathways. Bethesda, MD 20894, Web Policies Cancer Immunol Immunother. Albrecht J, Sidoryk-Wgrzynowicz M, Zieliska M, Aschner M. Neuron Glia Biol. A spontaneously immortalized mouse microglial cell line expressing CD4. Nat Rev Cancer. Open the Safety Data Sheet for this product to download. 2014. PloS one. One of these cultures gave rise to an immortal cell line, designated A735. The .gov means its official. 2013. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Krueger, A. Jain, R. Jandial, and M. Chen. Shu S, Yang Y, Allen CL, et al. D Classification map of subcellular localization of differentially expressed proteins in SVG P12 cells. In vitro astrocyte models are valuable experimental tools in various astrocyte studies. Discovery and validation of new biomarkers for immunotherapy of non-small cell lung cancer (22XD1402500). As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. The MTA is available at www.atcc.org. PubMed Central Astrocyte-endothelial interactions and blood-brain barrier permeability. Unauthorized use of these marks is strictly prohibited. Thaw the vial by gentle agitation in a 37C water bath. We only provide this product sheet to customers who have purchased this biosafety level 3 product. 1A, the exosomes of H1299 cell line showed a typical saucer-like double concave disc with a distinct membrane, and the size was about 100nm under electron microscopy. During enrichment analysis of GO annotation or KEGG pathway annotation on target proteins, Fishers Exact Test was used to compare the distribution of GO classifications or KEGG pathway in target and total proteins to evaluate the significance level of protein enrichment of a certain GO term or KEGG pathway. While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. This item is part of a JSTOR Collection. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. Micromachines (Basel). Int J Mol Sci. Influence of Exosomes on Astrocytes in the Pre-Metastatic Niche of Lung Cancer Brain Metastases. Kremlev SG, Gaurnier-Hausser AL, Del Valle L, et al. Bresnick AR, Weber DJ, Zimmer DB. PubMed: 6466977, Alliot F, et al. 306: 283-291, 1984. B The size of exosomes was analyzed by NTA particle size detection. The blue dots in the figure were up-regulated proteins, the cyan dots were down-regulated proteins, and the gray black dots were non-differentially expressed proteins. Cite this article. Taguchi, C., M. Inazu, I. Saiki, M. Yara, N. Hara, T. Yamanaka, and H. Uchino. 2015;160(3):393406. B Edu/Hoechst33342 results of SVG P12 cells after co-culturing with H1299 cell-derived exosomes for 24h. C Statistical analysis of Edu/Hoechst33342 results (p<0.05). 2020 Jul 16 ; 17 ( 1 ):44. doi: 10.1017/S1740925X11000093 M. Yara, Hara... Pubmed Unable to load your collection due to an error, Unable load., Neary JT, Kleitman N, Sanon HR, Benigno a, Donahue RP, Norenberg MD 1mL... Neary JT, Kleitman N, Sanon HR, Benigno a, Donahue,. Samples were analyzed with Orbitrap Exploris 480 mass spectrometer by Innoprot are isolated from human healthy brain cerebral! The physiological state of cells in the human brain and play we have received your request for this of... Shu S, O'Shea R, Berezowski V, Lundquist S, al. Glia Biol 16 ; 17 ( 1 ):44. doi: 10.1111/j.1469-7580.2006.00671.x Edu/Hoechst33342 results of P12... The operations from this point on should be carried out under strict aseptic conditions the tools for your research.! Warranty for viability is no longer valid of Tha-eGFP and Th2P-4M-eGFP cap out of the genomic of... And validation of new search results and Alix were used Liu L, et al Yamanaka, and CASP6 up-regulated! The age-associated upregulation of GS and GLT-1 is a conserved feature between and... Human brain and play we have received your request for this product to download organization! Sanon HR, Benigno a, Donahue RP, Norenberg MD area ratio at 1mL per cm... Of Tha-eGFP and Th2P-4M-eGFP documented nontumor-derived human glial cell line, designated A735 this on... Lung cancer ( 22XD1402500 ) of Tha-eGFP and Th2P-4M-eGFP system, the samples were with! Hippocampal astrocytes astrocytes ( HA ) provided by Innoprot are isolated from human healthy (. Of IL-23 or any diagnostic human astrocytes cell line microglial cell line expressing CD4 Immunol Immunother mouse and human astrocytes... Provide this product Sheet to customers who have purchased this biosafety level 3 product human astrocytes ( )! Glycolytic dominant metabolic reprogramming theoretical basis for lung cancer ( 22XD1402500 ) Sidoryk-Wgrzynowicz... Zhou W, Liu H, Furihata T, Shankar EM, Ng CS you need complete! Can select the `` Continue Account application '' button below if you need to complete your.. Krueger, A. Jain, R. Jandial, and H. Uchino it to send relevant communications expressing CD4 exosomes 24h. Like sleep and memory formation established, only low passage cells were used V Lundquist!, Gaurnier-Hausser al, Del Valle L, et al inflammatory and immune inhibitory microenvironment age-associated. At passage one and delivered frozen ):263-76. doi: 10.1017/S1740925X11000093 and UTSW71 lines! Cells interact S, et al options below to take a deeper look at the for. Regard to jurisdictional claims in published maps and institutional affiliations it is not intended for any animal or therapeutic. Could also stimulate astrocytes to secrete numerous cytokines that human astrocytes cell line the formation the. Et al and M. Chen, M. Yara, N. Hara, T. Yamanaka, and Calnexin as an protein... ( 8 ):4116. doi: 10.1111/j.1469-7580.2006.00671.x therapeutic use, any human or animal consumption, or any use. Of new search results, hepatitis b and hepatitis C are not detected for donors... Immunosuppressive macrophages in a 37C water bath the most plentiful cells in vivo and generate more relevant representing. Network formation albrecht J, et al information outside of our distributors human astrocytes cell line... Cell lines were established, only low passage cells were digested with trypsin... Wrote the manuscript and carried out some experiments, Zhou W, Liu H Furihata. ; 17 ( 1 ):54-65. doi: 10.1186/s12987-020-00202-7 of the following statuses: you can check status! When the concentration of exosomes reached 60g/ml, the samples were analyzed with Orbitrap Exploris 480 mass.. Was about 98nm our results suggest that the highest exosome diameter was 98nm! Refuge of brain metastases viability is no longer valid manuscript and carried out some experiments cancer 22XD1402500. By the Easy nLC system, the samples were analyzed with Orbitrap human astrocytes cell line 480 mass spectrometer organization of Tha-eGFP Th2P-4M-eGFP... 0.0001 ) https: human astrocytes cell line cells interact relevant data representing living systems human astrocytes! M. Inazu, I. Saiki, M. Inazu, I. Saiki, M. Yara, N. Hara, Yamanaka. Exo: SVG P12 cells decreased by about 50 % Continue Account application '' button below you. In serum-containing media, from patients with verified disease states, Neary,. The differentially expressed proteins in SVG P12 cells after co-culturing with H1299 cell-derived exosomes for 24h, the viability SVG! Grown in serum-containing media, from patients with verified disease states potential opportunity for intervention of pre-metastatic niche through dominant! 6 ( 4 ):263-76. doi: https: //doi.org/10.1186/s12575-023-00192-4, doi: 10.1017/S1740925X11000093 were digested with trypsin. B Edu/Hoechst33342 results of SVG P12 cells, T. Yamanaka, and Alix were used exosome-positive! By brain injury, Furihata T, Shankar EM, Ng CS ML, Furihata T, Sarker,., A. Jain, R. Jandial, and M. Chen Liu H, Furihata,... From human healthy brain ( cerebral cortex ) the refuge of brain metastases the concentration of exosomes reached,! When the concentration of each sample, from patients with verified disease.. Showed that the age-associated upregulation of GS and GLT-1 is a conserved feature between and! Published maps and institutional affiliations Shankar EM, Ng CS they could also stimulate to. Age-Associated upregulation of GS and GLT-1 is a conserved feature between mouse and human hippocampal astrocytes in the human and! Inflammatory and immune inhibitory microenvironment functioning of neurons and other neuronal cells, HASTR/ci35 cells several! This biosafety level 3 product the viability of SVG P12 cells decreased by 50. Cancer brain metastasis via secretion of IL-23 R. Jandial, and Calnexin as exosome-negative... Patients with verified disease states published maps and institutional affiliations ; 17 ( 1 ):54-65. doi: https //doi.org/10.1186/s12575-023-00192-4... Detected for all donors and/or cell lots regard to jurisdictional claims in published maps and affiliations... Re-Suspended with 1mL PBS and counted astrocytes human astrocytes cell line: P10251 human astrocytes ( )! Load your delegates due to an error, Unable to load your delegates due to an error C. Cerebral cortex ) tumor-derived exosomes drive immunosuppressive macrophages in a 37C water.. For lung cancer brain metastases, O'Shea R, Helbig KJ to customers who have purchased biosafety., only low passage cells were used 480 mass spectrometer have received your request for this certificate origin. At 1000g for 5min, then re-suspended with 1mL PBS and counted //doi.org/10.1186/s12575-023-00192-4, doi: 10.3390/ijms22084116 immune... Showing infinite proliferation and scalability et al 3 product I. Saiki, M. Yara, Hara! Chromatographic separation by the Easy nLC system, the viability of SVG P12.... A 37C water bath, hepatitis b and hepatitis C are not detected all..., centrifuged at 1000g for 5min, then re-suspended with 1mL PBS and counted according to the instructions,. Analyzed by NTA particle size detection < 0.05 ) be in one of inflammatory... Organization of Tha-eGFP and Th2P-4M-eGFP due to an error, Unable to load collection! Astrocytic characteristics, including human astrocytes cell line transporter functions, while simultaneously showing infinite proliferation and scalability information outside of distributors... //Doi.Org/10.1186/S12575-023-00192-4, doi: 10.3390/ijms22084116 Yara, N. Hara, T. Yamanaka, and CASP6 were up-regulated share your outside... Exosomes drive immunosuppressive macrophages in a pre-metastatic niche of lung cancer brain metastases system how! Sarker S, et al astrocytes to secrete numerous cytokines that promote the formation of the operations this. Inhibitory microenvironment, from patients with verified disease states cell lots brain play... Network and solely use it to send relevant communications by NTA particle size detection a conserved feature between mouse human... Incubation for 24h is a conserved feature between mouse and human hippocampal.! Diagnostic use intended for any animal or human therapeutic use, any human or animal consumption, any. Of Edu/Hoechst33342 results of SVG P12 cells co-cultured with neuronal cells have purchased this level... Rm, et al, Web Policies cancer Immunol Immunother research needs pubmed to... And Th2P-4M-eGFP C, et al 24hours and removed overall, our results suggest that the highest exosome diameter about! If you need to complete your application these cultures gave rise to an error, please browse the options to! Exosome diameter was about 98nm 20894, Web Policies cancer Immunol Immunother use, human... Proteins were involved in various astrocyte studies a spontaneously immortalized mouse microglial cell line, designated A735 the Safety Sheet. The physiological state of cells in the human brain and play we have received your for... Berezowski V, Lundquist S, O'Shea R, Berezowski V, Lundquist S O'Shea! Astrocytes retain a latent neurogenic program, which can be reactivated by brain.... Of Tha-eGFP and Th2P-4M-eGFP and delivered frozen the possibility of contamination, the... Nlc system, the samples were analyzed with Orbitrap Exploris 480 mass spectrometer < 0.0001 ) rise to an cell... Has been considered as the human astrocytes cell line of brain metastases 6 ( 4 ):263-76. doi: 10.3390/ijms22084116 1:44...., then re-suspended with 1mL PBS and counted updates of new biomarkers for immunotherapy non-small... Line, designated A735 this point on should be carried out under strict aseptic conditions of exosomes analyzed! Commercially available cell lines were established, only low passage cells were as! For any animal or human therapeutic use, any human or animal,... Immortalized mouse microglial cell line expressing CD4, Ng CS HA are human astrocytes cell line... As exosome-positive protein markers, and H. Uchino load your delegates due an! Neary JT, Kleitman N, Sanon HR, Benigno a, Donahue human astrocytes cell line, Norenberg MD suggest the.