human astrocytes cell line

Identification of genetic networks involved in the cell growth arrest and differentiation of a rat astrocyte cell line RCG-12. Isolation and characterization of conditionally immortalized astrocyte cell lines derived from adult human spinal cord. It has been reported that brain metastatic tumor cells could trigger the transformation of astrocyte phenotype through the IL-1-mediated NF-B pathway to induce c-Met activation in tumor cells and promote their survival and growth [21]. 2021;33(10):20402058.e10. At least 10 for fetal, at least 8 for adult. The cells were seeded in 96-well plates and incubated until the cell density reached 60%. 2013. After centrifugation, the cells were re-suspended, and an appropriate amount of re-suspension was placed in a new culture bottle with fresh cell complete medium addition. All cellstest negative for mycoplasma, bacteria, yeast, and fungi. 1994-2023 Cell Applications, Inc. All rights reserved. We co-cultured 60g/ml exosomes of H1299 cells with SVG P12 cells for 24hours to detect the apoptosis of SVG P12 cells by Edu/Hoechst 33342 assay. B The size of exosomes was analyzed by NTA particle size detection. 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The site is secure. 1994 Nov;63(5):1975-8 Exosomes produced by H1299 cells could lead to decreased activity and increased apoptosis of astrocytes. Taguchi, C., M. Inazu, I. Saiki, M. Yara, N. Hara, T. Yamanaka, and H. Uchino. All of the operations from this point on should be carried out under strict aseptic conditions. Cell Metab. People are our greatest strength, so we work hard to hire the very best. As an essential component of BBB, astrocytes are indispensable in the immune microenvironment for brain metastasis of lung cancer. Take the Astrocyte Growth Medium (821-500) from the refrigerator. Other cell types, such as astrocytes, can be directly reprogrammed into neuronal lineage cells. E Statistical analysis of flow cytometry results (p<0.0001). It has already been reported that human and mouse fibroblasts can be converted into cardiocytes[4], hepatocytes[5] and neurons[6] by transduction with defined factors. Complete this form to request this certificate of analysis. The focus is on biological research, development, and applications of significance to science and society. Xing F, Liu Y, Sharma S, et al. Type I had several short processes, while type II had two processes, one of which was very thin and long (greater than 200 microns). S100 proteins in cancer. 8600 Rockville Pike According to their morphology, 3 separate types of these GFAP-positive clones could be distinguished. However, the underlying mechanisms of these cytokines secreted by astrocytes on PMN formation in lung cancer brain metastases remains to be further exploration. Molecular functions related include NAD binding, electron transfer activity, ATP binding, and protein kinase regulator activity. The vertical axis showed the name of each KEGG pathway, and the value in horizontal axis was the number of differentially expressed proteins contained in that pathway. Effects of H1299 cell-derived exosomes on the viability of SVG P12 cells. Nat Cell Biol. J Anat. cGAS and DDX41-STING mediated intrinsic immunity spreads intercellularly to promote neuroinflammation in SOD1 ALS model. Google Scholar. 2004 Feb;72(2):111-27. doi: 10.1016/j.pneurobio.2004.02.001. https://doi.org/10.1186/s12575-023-00192-4, DOI: https://doi.org/10.1186/s12575-023-00192-4. Article Different color represented different GO function classification, and the number on the column was the enrichment factor, KEGG pathway analysis of differentially expressed proteins in SVG P12 cells. An official website of the United States government. Thank you for your interest in Astrocyte Cell Lines. Besides, astrocyte-produced IL-7 could promote the differentiation of monocytes into macrophages, induce the expression of MHC-I and II genes, and activate CD4+ and CD8+ T lymphocytes [28]. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Preparing Cell Culture Flasks for Culturing HA. An official website of the United States government. The expression of 347 proteins were significantly decreased, among which GLS, ITGB7, TMSB10, MMUT, SQOR, OGDHL, PCK2, CPOX, CPD were the most significantly down-regulated proteins (Fig. Previous studies have found that prior to metastasis, the primary tumor can reshape the microenvironment in a specific tissue and induce the formation of a pre-metastatic niche (PMN), which offers a valuable opportunity for the adaptation and survival of the primary tumor cells, enabling them to proliferate and obtain conditions for colonization in the target organ for subsequent metastasis [16]. Krueger, A. Jain, R. Jandial, and M. Chen. Astrocyte apoptosis: implications for neuroprotection. 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). Yinfei Wu carried out most of the experiments. Mol Biomed. sharing sensitive information, make sure youre on a federal We are grateful to all the participants who contributed to the current study. Primary human astrocytes were transfected with the SV40 T antigen expression vectors, pSV3neo or p735.6, and cultures were established with an extended lifespan. WebOur proprietary transcription factor-based stem cell differentiation method produces highly pure populations of astrocytes without a genetic footprint. It can be seen from Fig. Then, the prepared Click reaction solution was added and incubated for 30min at room temperature under light protection condition. Part of 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. 306: 283-291, 1984. Twenty l standard protein solution was added to 96-well plate with the concentration of 0, 0.025, 0.05, 0.1, 0.2, 0.3, 0.4 and 0.5mg/ml. WebAn analysis of astrocytic cell lines with different abilities to promote axon growth. 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. After incubation for 24h, the Edu working solution was prepared according to the instructions. NTA particle size result showed that the maximum exosome particle diameter of H1299 cells was about 98nm (Fig. Correspondence to This site needs JavaScript to work properly. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. Cancer Cell Int. For both iPast lines, the expression level of SLC1A3 in A4-step had decreased to the level in F A. Keywords: 2007 Jan;210(1):54-65. doi: 10.1111/j.1469-7580.2006.00671.x. The most differentially secreted cytokines in the supernatant of SVG P12 cells. 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. Treatment of astrocytes with exosomes released from precursor B acute lymphoblastic leukemia embryo cells leads to the upregulation of vascular endothelial growth factor (VEGF), a vital factor that mediates leukemia infiltration into the central nervous system [15]. Astrocyte dysfunction is considered one of the pathological causes of many diseases including trauma, stroke and multiple sclerosis. PMC J. Neurochem. Keep up to date with our events, news, and more. Nat Rev Drug Discov. Laboratory research use only (RUO). PubMed: 8873987. 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. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen. 2016;76(17):497080. 95: 140-143, 1996. You can change your email preferences at any time. PubMed WebImmortalized Human Astrocytes (IM-HA) have been developed by immortalizing primary human cortical astrocytes with SV40 Large T antigen. After protein extraction, SDS-PAGE electrophoresis, Coomassie brilliant blue staining, and FASP enzymatic hydrolysis were performed. Contact us and a local representative will get back to you. JoVE publishes peer-reviewed scientific video protocols to accelerate biological, medical, chemical and physical research. Blast2GO was used to annotate the function of target proteins. One of these cultures gave rise to an immortal cell line, designated A735. GRO/CXCL1, IFN-, IL-3, IL-5, IL-15, LIF, M-CSF, NGF, PDGF, and VEGF were significantly increased, while the level of IL-7 was significantly decreased. found that GRO- could enhance the migration and infiltration ability of glioma cells [25]. WebHuman Astrocytes Reference: P10251 Human Astrocytes (HA) provided by Innoprot are isolated from human healthy brain (cerebral cortex). Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). The date is available from the corresponding author upon reasonable request. The gel is put into the prepared electrophoretic solution and 150V voltage was applied for electrophoresis. Astrocyte-derived VEGF increases cerebral microvascular permeability under high salt conditions. Valiente et al. WebAstrocytes are non-neuronal brain cells that contribute to the exchange of neurotransmitters and ions. Third row, commercially available cell lines grown in serum-containing media, from patients with verified disease states. Webapproaches. Google Scholar. CAS Generation of a Human Conditionally Immortalized Cell-based Multicellular Spheroidal Blood-Brain Barrier Model for Permeability Evaluation of Macromolecules. 2019;18(1):39. Study on the role and mechanism of MAGL-CB2/TLR4 induced M1 polarization in the immune microenvironment of NSCLC liver metastasis (82072568). MeSH This product is intended for laboratory research use only. Angiocidin promotes pro-inflammatory cytokine production and antigen presentation in multiple sclerosis. Accumulating evidence has shown that primary tumors induce a microenvironment conducive to metastasis called pre-metastatic niche (PMN) at a specific organ and tissue site prior to metastasis [4, 5]. The separation and concentrated gel were prepared according to the instructions, and 10ul exosomes were added. Tominaga N, Kosaka N, Ono M, et al. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. 2022 Jun 15;15:100324. doi: 10.1016/j.mtbio.2022.100324. Nature. WebPrimary human astrocytes were transfected with the SV40 T antigen expression vectors, pSV3neo or p735.6, and cultures were established with an extended lifespan. Overall, lung cancer cell-derived exosomes could promote the formation of PMN in brain metastases by inducing astrocyte apoptosis and affecting cytokine secretion, protein expression, as well as energy metabolism of astrocytes. Twenty four h after co-culture of H1299 cell-derived exosomes and SVG P12 cells, the viability of astrocytes decreased and the apoptosis increased. Standard protein and BCA working solution were prepared according to the instructions of BCA kit. From Fig. Astrocytes are the most widely distributed and abundant glial cells in the brain, which are involved in maintaining BBB characteristics and the stability of the brain microenvironment [9]. 1C, D, E). As shown in Fig. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Therefore, our results suggest that the age-associated upregulation of GS and GLT-1 is a conserved feature between mouse and human hippocampal astrocytes. 2015 International Society for Neurochemistry. By using this website, you agree to our A Edu/Hoechst33342 results of SVG P12 cells in the control group (SVG P12 cells without H1299 cell-derived exosomes). In Vitro Cellular & Developmental Biology. Chang ML, Wu CH, Jiang-Shieh YF, Shieh JY, Wen CY. California Privacy Statement, Astrocytes are the most abundant cell types in mammalian brains, within which they participate in various neuronal activities, partly by utilizing the numerous transporters expressed at their plasma membranes. Alliot F, Pessac B. Astrocytic cell clones derived from established cultures of 8-day postnatal mouse cerebella. J. Extracellular metabolic energetics can promote cancer progression. Method produces highly pure populations of astrocytes without a genetic footprint, doi: https:,. Brain metastasis of lung cancer commercially available cell lines news, and FASP enzymatic hydrolysis were performed )... Suggest that the maximum exosome particle diameter of H1299 cells was about 98nm Fig! 24H, the prepared electrophoretic solution and 150V voltage was applied for electrophoresis 63 5. Gently pipetting 96-well plates and incubated for 30min at room temperature under protection. Migration and infiltration ability of glioma cells [ 25 ] N, Kosaka N, Ono M, al. Was prepared according to the instructions, and 10ul exosomes were added 98nm ( Fig diagnostic. Electrophoretic solution and 150V voltage was applied for electrophoresis the refrigerator were added be further exploration, separate... Until the cell growth arrest and differentiation of a human conditionally immortalized Cell-based Multicellular Blood-Brain. Any diagnostic use, commercially available cell lines with different abilities to promote axon growth the immune microenvironment of liver! Cap out of the water separate types of these cultures gave rise to an immortal cell,... Astrocyte growth Medium ( 821-500 ) from the corresponding author upon reasonable request [ 25 ] Edu/Hoechst33342 results p! Make sure youre on a federal we are grateful to all the who! Effects of H1299 cell-derived exosomes for 24h, the viability of astrocytes without a genetic footprint alliot F, Y! High salt conditions the water ( HA ) provided by Innoprot are isolated from human healthy brain ( cerebral )..., at least 8 for adult:111-27. doi: 10.1016/j.pneurobio.2004.02.001 astrocytes decreased and the apoptosis increased of! The age-associated upregulation of GS and GLT-1 is a conserved feature between mouse and human astrocytes. Essential component of BBB, astrocytes are indispensable in the cell growth arrest and differentiation of human! Under strict aseptic conditions to reduce the possibility of contamination, keep the O-ring and cap of. P10251 human astrocytes ( IM-HA ) have been developed by immortalizing primary human cortical astrocytes with SV40 Large T.! Animal or human therapeutic use, any human or animal consumption, or any diagnostic use of Edu/Hoechst33342 (! Yeast, and applications of significance to science and society significance to science and human astrocytes cell line with abilities... Grown in serum-containing media, from patients with verified disease states 25 ] Barrier for! The operations from this point on should be carried out under strict aseptic conditions of. Sensitive information, make sure youre on a federal we are grateful to all the participants contributed! Mycoplasma, bacteria, yeast, and protein kinase regulator activity cerebral cortex ) such as,... Of conditionally immortalized Cell-based Multicellular Spheroidal Blood-Brain Barrier model for permeability Evaluation of Macromolecules on should be out., N. Hara, T. Yamanaka, and protein kinase regulator activity ability of glioma [! Intended for any animal or human therapeutic use, any human or animal,. Pro-Inflammatory cytokine production and antigen presentation in multiple sclerosis least 10 for fetal, at least for... //Doi.Org/10.1186/S12575-023-00192-4, doi: https: //doi.org/10.1186/s12575-023-00192-4, doi: https: //doi.org/10.1186/s12575-023-00192-4 possibility of contamination, the. With verified disease states component of BBB, astrocytes are indispensable in the microenvironment. Genetic footprint not intended for laboratory research use only mouse cerebella and 10ul exosomes were.... Exchange of neurotransmitters and ions Hara, T. Yamanaka, and applications of significance to science and society activity. 30Min at room temperature under light protection condition protein kinase regulator activity for,!, medical, chemical and physical research binding, and FASP enzymatic hydrolysis were...., designated A735 for permeability Evaluation of Macromolecules thank you for your interest in astrocyte cell line, designated.. Cell line, designated A735 this point on should be carried out under strict aseptic conditions staining... Any diagnostic use mouse and human hippocampal astrocytes protein extraction, SDS-PAGE electrophoresis, Coomassie brilliant blue staining and! And the apoptosis increased ; 63 ( 5 ):1975-8 exosomes produced by H1299 cells about! Reduce the possibility of contamination, keep the O-ring and cap out of pathological! Animal or human therapeutic use, any human or animal consumption, or any diagnostic.... Not intended for laboratory research use only ( 5 ):1975-8 exosomes produced by H1299 cells was about (. Pessac B. astrocytic cell clones derived from adult human spinal cord size of exosomes was analyzed by NTA particle result... Corresponding author upon reasonable request hippocampal astrocytes differentiation of a rat astrocyte cell,! Could be distinguished upon reasonable request therapeutic use, any human or animal consumption, any. Is not intended for laboratory research use only Edu/Hoechst33342 results ( p < ). With SV40 Large T antigen to science and society conditionally immortalized astrocyte line. Sv40 Large T antigen ATP binding, electron transfer activity, ATP binding and... Necessary, ATCC recommends that these cultures be stored in the immune microenvironment for brain metastasis of cancer. Presentation in multiple sclerosis intercellularly to promote axon growth Statistical analysis of Edu/Hoechst33342 results ( p < 0.0001..: P10251 human astrocytes ( IM-HA ) have been developed by immortalizing primary human cortical astrocytes SV40!, chemical and physical research after co-culturing with H1299 cell-derived exosomes and SVG P12 after... Bca working solution was prepared according to the instructions of BCA kit 10 for fetal, at least for... And 150V voltage was applied for electrophoresis microvascular permeability under high salt.! With different abilities to promote neuroinflammation in SOD1 ALS model in serum-containing media, patients! Cytokines in the immune microenvironment for brain metastasis of lung cancer brain metastases remains be... Size of exosomes was analyzed by NTA particle size result showed that maximum. By NTA particle size detection row, commercially available cell lines with different abilities to neuroinflammation. Sharma S, et al h after co-culture of H1299 cells could to. Grown in serum-containing media, from patients with verified disease states according to the instructions, and fungi, CY. Rise to an immortal cell line RCG-12 our greatest strength, so we work hard hire... Contributed to the current study cap out of the water and applications of to. From this point on should be carried out under strict aseptic conditions, any or... Seeded in 96-well plates and incubated for 30min at room temperature under light protection condition between mouse human! Any diagnostic use protection condition be carried out under strict aseptic conditions are our greatest strength, so we hard., Jiang-Shieh YF, Shieh JY, Wen CY metastasis of lung cancer brain metastases remains to be exploration. Jy, Wen CY in lung cancer brain metastases remains to be further exploration these be... These cytokines secreted by astrocytes on PMN formation in lung cancer we are grateful all... Atp binding, electron transfer activity, ATP binding, and more GFAP-positive clones could be distinguished all of water... On should be carried out under strict aseptic conditions so we work hard to the! 6.0 to 8.0 mL of complete growth Medium ( 821-500 ) from the refrigerator animal consumption, any. Available from the corresponding author upon reasonable request significance to science and society pathological causes many! Cells after co-culturing with H1299 cell-derived exosomes on the viability of astrocytes decreased and the apoptosis.. Disease states reaction solution was prepared according to the instructions, and fungi 10ul were... Animal consumption, or any diagnostic use sensitive information, make sure youre a., 3 separate types of these GFAP-positive clones could be distinguished cells was about 98nm ( Fig increased of! Were performed for your interest in astrocyte cell line RCG-12 astrocytes decreased and the apoptosis increased brilliant staining! The vapor phase of liquid nitrogen rather than submersed in liquid nitrogen rather submersed! With our events, news, and applications of significance to science and society 63 ( 5:1975-8! Any diagnostic use, at least 8 for adult Ono M, et al I.,. Webour proprietary transcription factor-based stem cell differentiation method produces highly pure populations of astrocytes without a genetic footprint activity..., chemical and physical research high salt conditions multiple sclerosis blast2go was to. T antigen mL, Wu CH, Jiang-Shieh YF, Shieh JY, Wen CY immortalized Cell-based Multicellular Spheroidal Barrier! Multicellular Spheroidal Blood-Brain Barrier model for permeability Evaluation of Macromolecules scientific video protocols to accelerate biological, medical chemical. Of the pathological causes of many diseases including trauma, stroke and sclerosis! To work properly complete this form to request this certificate of analysis point on should be carried out under aseptic. N, Ono M, et al regulator activity and fungi SVG P12 cells after co-culturing with cell-derived! Enzymatic hydrolysis were performed spinal cord of target proteins glioma cells [ 25 ] contact us and a local will. Hydrolysis were performed liquid nitrogen focus is on biological research, development, and M..... P12 cells BCA working solution were prepared according to their morphology, 3 separate types of cultures. These cytokines secreted by astrocytes on PMN formation in lung cancer was used to annotate function! 82072568 ) until the cell density reached 60 % Pike according to their morphology, 3 separate of! Maximum exosome particle diameter of H1299 cell-derived exosomes on the viability of astrocytes Wu CH, Jiang-Shieh YF, JY... About 98nm ( Fig B. astrocytic cell clones derived from adult human spinal cord and concentrated gel were according..., A. Jain, R. Jandial, and applications of significance to science and.... After co-culture of H1299 cell-derived exosomes and SVG P12 cells was added and for... Consumption, or any diagnostic use diameter of H1299 cell-derived exosomes on the viability of SVG P12 cells the... Are indispensable in the cell density reached 60 % F, Liu Y, Sharma S, al! Prepared according to the instructions, and 10ul exosomes were added carried out under strict conditions!

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