Name: CD63-GFP Raporlayıcı Fare Modeli Kullanılarak Adipositten Türetilen Ekstraselüler Vezikül Salgısının Karakterizasyonu In Vivo ve In Vitro (Video) | JoVE Journal
Uploaded: 2025-12-05T13:20:00.000+00:00
Duration: 10 min 40 s
Description: Keşfedin, CD63-GFP raporlayıcı fare modeli kullanarak adiposit kaynaklı ekstraselüler vezikül salgısını in vivo ve in vitro karakterize etme yöntemini. Bu protokol metabolik düzenleme ve hücre iletişimi araştırmaları için optimize edilmiş bir sistem sunar.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

<ol><li>Kershaw, E. E., Flier, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipose+tissue+as+an+endocrine+organ%5BTitle%5D)+AND+%22J+Clin+Endocrinol+Metab%22%5BJournal%5D)">Adipose tissue as an endocrine organ.</a> J Clin Endocrinol Metab. 89 (3), 2548-2556 (2004).</li>
<li>Wu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Beige+adipocytes+are+a+distinct+type+of+thermogenic+fat+cell+in+mouse+and+human%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human.</a> Cell. 150 (2), 366-376 (2012).</li>
<li>Luo, L., Liu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipose+tissue+in+control+of+metabolism%5BTitle%5D)+AND+%22J+Endocrinol%22%5BJournal%5D)">Adipose tissue in control of metabolism.</a> J Endocrinol. 231 (3), R77-R99 (2016).</li>
<li>Lynes, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+cold-induced+lipokine+12%2C13-diHOME+promotes+fatty+acid+transport+into+brown+adipose+tissue%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">The cold-induced lipokine 12,13-diHOME promotes fatty acid transport into brown adipose tissue.</a> Nat Med. 23, 631-637 (2017).</li>
<li>Sakers, A., De Siqueira, M. K., Seale, P., Villanueva, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipose-tissue+plasticity+in+health+and+disease%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Adipose-tissue plasticity in health and disease.</a> Cell. 185 (3), 419-446 (2022).</li>
<li>Santoro, A., McGraw, T. E., Kahn, B. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Insulin+action+in+adipocytes%2C+adipose+remodeling%2C+and+systemic+effects%5BTitle%5D)+AND+%22Cell+Metab%22%5BJournal%5D)">Insulin action in adipocytes, adipose remodeling, and systemic effects.</a> Cell Metab. 33 (4), 748-757 (2021).</li>
<li>Czech, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanisms+of+insulin+resistance+related+to+white%2C+beige%2C+and+brown+adipocytes%5BTitle%5D)+AND+%22Mol+Metab%22%5BJournal%5D)">Mechanisms of insulin resistance related to white, beige, and brown adipocytes.</a> Mol Metab. 34, 27-42 (2020).</li>
<li>Kahn, S. E., Hull, R. L., Utzschneider, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanisms+linking+obesity+to+insulin+resistance+and+type+2+diabetes%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Mechanisms linking obesity to insulin resistance and type 2 diabetes.</a> Nature. 444, 840-846 (2006).</li>
<li>Feng, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SNAP23+regulates+BAX-dependent+adipocyte+programmed+cell+death+independently+of+canonical+macroautophagy%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">SNAP23 regulates BAX-dependent adipocyte programmed cell death independently of canonical macroautophagy.</a> J Clin Invest. 128 (9), 3941-3956 (2018).</li>
<li>Ouchi, N., Parker, J. L., Lugus, J. J., Walsh, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipokines+in+inflammation+and+metabolic+disease%5BTitle%5D)+AND+%22Nat+Rev+Immunol%22%5BJournal%5D)">Adipokines in inflammation and metabolic disease.</a> Nat Rev Immunol. 11, 85-97 (2011).</li>
<li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Positional+cloning+of+the+mouse+obese+gene+and+its+human+homologue%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Positional cloning of the mouse obese gene and its human homologue.</a> Nature. 372, 425-432 (1994).</li>
<li>Scherer, P. E., Williams, S., Fogliano, M., Baldini, G., Lodish, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+serum+protein+similar+to+C1q%2C+produced+exclusively+in+adipocytes%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">A novel serum protein similar to C1q, produced exclusively in adipocytes.</a> J Biol Chem. 270 (45), 26746-26749 (1995).</li>
<li>Considine, R. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Serum+immunoreactive-leptin+concentrations+in+normal-weight+and+obese+humans%5BTitle%5D)+AND+%22N+Engl+J+Med%22%5BJournal%5D)">Serum immunoreactive-leptin concentrations in normal-weight and obese humans.</a> N Engl J Med. 334, 292-295 (1996).</li>
<li>Hotamisligil, G. S., Shargill, N. S., Spiegelman, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipose+expression+of+tumor+necrosis+factor-alpha%3A+Direct+role+in+obesity-linked+insulin+resistance%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Adipose expression of tumor necrosis factor-alpha: Direct role in obesity-linked insulin resistance.</a> Science. 259 (5091), 87-91 (1993).</li>
<li>Steppan, C. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+hormone+resistin+links+obesity+to+diabetes%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">The hormone resistin links obesity to diabetes.</a> Nature. 409, 307-312 (2001).</li>
<li>Yang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Serum+retinol+binding+protein+4+contributes+to+insulin+resistance+in+obesity+and+type+2+diabetes%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Serum retinol binding protein 4 contributes to insulin resistance in obesity and type 2 diabetes.</a> Nature. 436, 356-362 (2005).</li>
<li>Tilg, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipokines%3A+Masterminds+of+metabolic+inflammation%5BTitle%5D)+AND+%22Nat+Rev+Immunol%22%5BJournal%5D)">Adipokines: Masterminds of metabolic inflammation.</a> Nat Rev Immunol. 25, 250-265 (2025).</li>
<li>Clemente-Suarez, V. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+role+of+adipokines+in+health+and+disease%5BTitle%5D)+AND+%22Biomedicines%22%5BJournal%5D)">The role of adipokines in health and disease.</a> Biomedicines. 11 (5), 1290(2023).</li>
<li>Hallal, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Understanding+the+extracellular+vesicle+surface+for+clinical+molecular+biology%5BTitle%5D)+AND+%22J+Extracell+Vesicles%22%5BJournal%5D)">Understanding the extracellular vesicle surface for clinical molecular biology.</a> J Extracell Vesicles. 11 (10), e12260(2022).</li>
<li>Welsh, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Minimal+information+for+studies+of+extracellular+vesicles+%28MISEV2023%29%3A+From+basic+to+advanced+approaches%5BTitle%5D)+AND+%22J+Extracell+Vesicles%22%5BJournal%5D)">Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches.</a> J Extracell Vesicles. 13 (2), e12404(2024).</li>
<li>Le Lay, S., Scherer, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exploring+adipose+tissue-derived+extracellular+vesicles+in+inter-organ+crosstalk%3A+Implications+for+metabolic+regulation+and+adipose+tissue+function%5BTitle%5D)+AND+%22Cell+Rep%22%5BJournal%5D)">Exploring adipose tissue-derived extracellular vesicles in inter-organ crosstalk: Implications for metabolic regulation and adipose tissue function.</a> Cell Rep. 44 (6), 115732(2025).</li>
<li>Shimoda, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exosome+surface+glycans+reflect+osteogenic+differentiation+of+mesenchymal+stem+cells%3A+Profiling+by+an+evanescent+field+fluorescence-assisted+lectin+array+system%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Exosome surface glycans reflect osteogenic differentiation of mesenchymal stem cells: Profiling by an evanescent field fluorescence-assisted lectin array system.</a> Sci Rep. 9, 11497(2019).</li>
<li>Albuquerque, P. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+Paracoccidioides+brasiliensis+glycan+shares+serologic+and+functional+properties+with+cryptococcal+glucuronoxylomannan%5BTitle%5D)+AND+%22Fungal+Genet+Biol%22%5BJournal%5D)">A Paracoccidioides brasiliensis glycan shares serologic and functional properties with cryptococcal glucuronoxylomannan.</a> Fungal Genet Biol. 49 (11), 943-954 (2012).</li>
<li>Li, Y., Tang, X., Gu, Y., Zhou, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipocyte-derived+extracellular+vesicles%3A+Small+vesicles+with+big+impact%5BTitle%5D)+AND+%22Front+Biosci+%28Landmark+Ed%29%22%5BJournal%5D)">Adipocyte-derived extracellular vesicles: Small vesicles with big impact.</a> Front Biosci (Landmark Ed). 28 (7), 149(2023).</li>
<li>Thomou, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipose-derived+circulating+miRNAs+regulate+gene+expression+in+other+tissues%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Adipose-derived circulating miRNAs regulate gene expression in other tissues.</a> Nature. 542, 450-455 (2017).</li>
<li>Crewe, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+endothelial-to-adipocyte+extracellular+vesicle+axis+governed+by+metabolic+state%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">An endothelial-to-adipocyte extracellular vesicle axis governed by metabolic state.</a> Cell. 175 (3), 695-708.e13 (2018).</li>
<li>Flaherty, S. E. 3rd, et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+lipase-independent+pathway+of+lipid+release+and+immune+modulation+by+adipocytes%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">A lipase-independent pathway of lipid release and immune modulation by adipocytes.</a> Science. 363 (6430), 989-993 (2019).</li>
<li>Clement, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipocyte+extracellular+vesicles+carry+enzymes+and+fatty+acids+that+stimulate+mitochondrial+metabolism+and+remodeling+in+tumor+cells%5BTitle%5D)+AND+%22EMBO+J%22%5BJournal%5D)">Adipocyte extracellular vesicles carry enzymes and fatty acids that stimulate mitochondrial metabolism and remodeling in tumor cells.</a> EMBO J. 39, e102525(2020).</li>
<li>Kulaj, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipocyte-derived+extracellular+vesicles+increase+insulin+secretion+through+transport+of+insulinotropic+protein+cargo%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Adipocyte-derived extracellular vesicles increase insulin secretion through transport of insulinotropic protein cargo.</a> Nat Commun. 14, 709(2023).</li>
<li>McCann, J. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reporter+mice+for+isolating+and+auditing+cell+type-specific+extracellular+vesicles+in+vivo%5BTitle%5D)+AND+%22Genesis%22%5BJournal%5D)">Reporter mice for isolating and auditing cell type-specific extracellular vesicles in vivo.</a> Genesis. 58 (7), e23369(2020).</li>
<li>Men, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exosome+reporter+mice+reveal+the+involvement+of+exosomes+in+mediating+neuron+to+astroglia+communication+in+the+CNS%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Exosome reporter mice reveal the involvement of exosomes in mediating neuron to astroglia communication in the CNS.</a> Nat Commun. 10, 4136(2019).</li>
<li>Jeffery, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+of+Cre+recombinase+models+for+the+study+of+adipose+tissue%5BTitle%5D)+AND+%22Adipocyte%22%5BJournal%5D)">Characterization of Cre recombinase models for the study of adipose tissue.</a> Adipocyte. 3 (3), 206-211 (2014).</li>
<li>Emont, M. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+single-cell+atlas+of+human+and+mouse+white+adipose+tissue%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">A single-cell atlas of human and mouse white adipose tissue.</a> Nature. 603, 926-933 (2022).</li>
<li>Krylova, S. V., Feng, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+machinery+of+exosomes%3A+Biogenesis%2C+release%2C+and+uptake%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">The machinery of exosomes: Biogenesis, release, and uptake.</a> Int J Mol Sci. 24 (2), (2023).</li>
<li>Crewe, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+challenges+of+interrogating+adipose+tissue+extracellular+vesicle+functions+in+physiology%5BTitle%5D)+AND+%22Commun+Biol%22%5BJournal%5D)">The challenges of interrogating adipose tissue extracellular vesicle functions in physiology.</a> Commun Biol. 5, 581(2022).</li>
<li>Blandin, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lipidomic+analysis+of+adipose-derived+extracellular+vesicles+reveals+specific+EV+lipid+sorting+informative+of+the+obesity+metabolic+state%5BTitle%5D)+AND+%22Cell+Rep%22%5BJournal%5D)">Lipidomic analysis of adipose-derived extracellular vesicles reveals specific EV lipid sorting informative of the obesity metabolic state.</a> Cell Rep. 42 (3), 112169(2023).</li>
<li>Matilainen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Increased+secretion+of+adipocyte-derived+extracellular+vesicles+is+associated+with+adipose+tissue+inflammation+and+the+mobilization+of+excess+lipid+in+human+obesity%5BTitle%5D)+AND+%22J+Transl+Med%22%5BJournal%5D)">Increased secretion of adipocyte-derived extracellular vesicles is associated with adipose tissue inflammation and the mobilization of excess lipid in human obesity.</a> J Transl Med. 22, 623(2024).</li>
<li>Thery, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Minimal+information+for+studies+of+extracellular+vesicles+2018+%28MISEV2018%29%3A+A+position+statement+of+the+International+Society+for+Extracellular+Vesicles+and+update+of+the+MISEV2014+guidelines%5BTitle%5D)+AND+%22J+Extracell+Vesicles%22%5BJournal%5D)">Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines.</a> J Extracell Vesicles. 7 (1), 1535750(2018).</li>
<li>Olson, A. L., Knight, J. B., Pessin, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Syntaxin+4%2C+VAMP2%2C+and%2For+VAMP3%2Fcellubrevin+are+functional+target+membrane+and+vesicle+SNAP+receptors+for+insulin-stimulated+GLUT4+translocation+in+adipocytes%5BTitle%5D)+AND+%22Mol+Cell+Biol%22%5BJournal%5D)">Syntaxin 4, VAMP2, and/or VAMP3/cellubrevin are functional target membrane and vesicle SNAP receptors for insulin-stimulated GLUT4 translocation in adipocytes.</a> Mol Cell Biol. 17 (5), 2425-2435 (1997).</li>
<li>Oh, E., Miller, R. A., Thurmond, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Syntaxin+4+overexpression+ameliorates+effects+of+aging+and+high-fat+diet+on+glucose+control+and+extends+lifespan%5BTitle%5D)+AND+%22Cell+Metab%22%5BJournal%5D)">Syntaxin 4 overexpression ameliorates effects of aging and high-fat diet on glucose control and extends lifespan.</a> Cell Metab. 22 (3), 499-507 (2015).</li>
<li>Merz, K. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Enrichment+of+the+exocytosis+protein+STX4+in+skeletal+muscle+remediates+peripheral+insulin+resistance+and+alters+mitochondrial+dynamics+via+Drp1%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Enrichment of the exocytosis protein STX4 in skeletal muscle remediates peripheral insulin resistance and alters mitochondrial dynamics via Drp1.</a> Nat Commun. 13, 424(2022).</li>
<li>Yu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Involution+of+brown+adipose+tissue+through+a+syntaxin+4+dependent+pyroptosis+pathway%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Involution of brown adipose tissue through a syntaxin 4 dependent pyroptosis pathway.</a> Nat Commun. 15, 2856(2024).</li>
<li>Verweij, F. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Quantifying+exosome+secretion+from+single+cells+reveals+a+modulatory+role+for+GPCR+signaling%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling.</a> J Cell Biol. 217 (3), 1129-1142 (2018).</li>
<li>Arora, A., Sharma, V., Gupta, R., Aggarwal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+and+characterization+of+extracellular+vesicles+derived+from+ex+vivo+culture+of+visceral+adipose+tissue%5BTitle%5D)+AND+%22Bio+Protoc%22%5BJournal%5D)">Isolation and characterization of extracellular vesicles derived from ex vivo culture of visceral adipose tissue.</a> Bio Protoc. 14 (11), e5011(2024).</li>
<li>Benedikter, B. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ultrafiltration+combined+with+size+exclusion+chromatography+efficiently+isolates+extracellular+vesicles+from+cell+culture+media+for+compositional+and+functional+studies%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Ultrafiltration combined with size exclusion chromatography efficiently isolates extracellular vesicles from cell culture media for compositional and functional studies.</a> Sci Rep. 7, 15297(2017).</li>
<li>Nordin, J. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ultrafiltration+with+size-exclusion+liquid+chromatography+for+high+yield+isolation+of+extracellular+vesicles+preserving+intact+biophysical+and+functional+properties%5BTitle%5D)+AND+%22Nanomedicine%22%5BJournal%5D)">Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties.</a> Nanomedicine. 11 (4), 879-883 (2015).</li>
</ol>

<table><tbody><tr><td>0.5 M EDTA, pH 8.0</td><td>Fisher</td><td>AM9260G</td><td>500x seyreltme</td></tr><tr><td>10x PBS</td><td>Fisher Scientific</td><td>70-013-032</td><td>1000x suyla seyreltme</td></tr><tr><td>1 M HEPES</td><td>Gibco</td><td>15630080</td><td>1000x seyrelme</td></tr><tr><td>Yağ Dokusu Progenitor İzolasyon Kiti, Fare</td><td>Miltenyi biyotec</td><td>30-106-639</td><td>Kullanıma hazır</td></tr><tr><td>Amicon Ultra Santrifüj Filtre, 100 kDa MWCO</td><td>Sigma</td><td>UFC9100</td><td>15 mL örnek hacmi</td></tr><tr><td>Amicon Ultra Santrifüj Filtre, 100 kDa MWCO</td><td>Sigma</td><td>UFC9100</td><td>4 mL örnek hacmi</td></tr><tr><td>BSA</td><td>Sigma</td><td>A6003</td><td>Kullanıma hazır</td></tr><tr><td>Kalneksin antikoru</td><td>calnexin</td><td>AB22595</td><td>1000x seyrelme</td></tr><tr><td>CD63 antikoru</td><td>abcam</td><td>AB217345</td><td>500x seyreltme</td></tr><tr><td>CD81</td><td>hücre sinyal teknolojisi</td><td>10037'ler</td><td>500x seyreltme</td></tr><tr><td>Corning&nbsp; BioCoat&nbsp; Collagen I Kültür Slaytı</td><td>Sigma</td><td>CLS354630</td><td>Kullanıma hazır</td></tr><tr><td>Corning&nbsp; Hücre süzcü</td><td>Sigma</td><td>CLS431750</td><td>40 &mu; m gözenek boyutu</td></tr><tr><td>Corning&nbsp; Hücre süzcü</td><td>Sigma</td><td>CLS431752</td><td>100 &mu; m gözenek boyutu</td></tr><tr><td>COX IV antikor</td><td>Hücre Sinyal Teknolojisi</td><td>4844</td><td>1000x seyrelme</td></tr><tr><td>DMEM</td><td>Gibco</td><td>11965092</td><td>Kullanıma hazır</td></tr><tr><td>DMEM/F-12, GlutaMAX takviyesi</td><td>Termobalıkçı</td><td>10565018</td><td>Kullanıma hazır</td></tr><tr><td>Dnase 1</td><td>Sigma</td><td>D4513</td><td>Stok 5 KU/mL -20 derece ve derece; C</td></tr><tr><td>GlutaMAX Takviyesi</td><td>Gibco</td><td>35050061</td><td>100x seyreltme</td></tr><tr><td>Halt&trade; Proteaz ve Fosfataz İnhibitörü Kokteyli (100X)</td><td>Termobalıkçı</td><td>78444</td><td>Kullanıma hazır</td></tr><tr><td>HSP70 antikoru</td><td>abcam</td><td>AB181606</td><td>1000x seyrelme</td></tr><tr><td>Invitrogen eBioscience 10X RBC Lizis Tamponu (Çok Türlü)</td><td>Fisher Scientific</td><td>50-112-9743</td><td>Suda 10 kez seyreltme</td></tr><tr><td>Izon Science Usa Ltd QEVORIGINAL 70NMGEN2 5/PK SDP</td><td>Fisher Scientific</td><td>NC2042225</td><td>Kullanıma hazır</td></tr><tr><td>LEICA STELLARIS 8 konfokal mikroskobu</td><td>Leica Mikroskopları</td><td>Yok</td><td></td></tr><tr><td>Liberase TM Araştırma Derecesi</td><td>Sigma</td><td>LIBTM-RO</td><td>Stok 5 mg/mL -20 derece ve derece; C</td></tr><tr><td>Millex MCE şırınga filtresi</td><td>Sigma</td><td>SLAA025</td><td>0.8 &mu; m gözenek boyutu</td></tr><tr><td>NanoSight Pro</td><td>Malvern Panalytical</td><td>Yok</td><td></td></tr><tr><td>Novex Tris-Glycine Mini Protein Jelleri, 4&ndash; 12%,</td><td>Termobalıkçı</td><td>XP04120BOX</td><td>Kullanıma hazır</td></tr><tr><td>Novex Tris-Glycine SDS Numune Tamponu (2x)</td><td>Termobalıkçı</td><td>LC2676</td><td>Kullanıma hazır</td></tr><tr><td>Pacific Blue anti-fare Ly-6A/E (Sca-1) Antikor</td><td>BioLegend</td><td>108119</td><td>25x seyreltme</td></tr><tr><td>Paraformaldehit çözeltisi (%16)</td><td>Fisher Scientific</td><td>50-980-487</td><td>PBS ile %4'e kadar seyreltin</td></tr><tr><td>pluriStrainer S / 30 &micro; m (Hücre Süzgeci)</td><td>Fisher Scientific</td><td>NC0922459</td><td>30 &mu; m gözenek boyutu</td></tr><tr><td>Ponceau S çözümü</td><td>Sigma</td><td>P7170-1L</td><td>Kullanıma hazır</td></tr><tr><td>Primocin</td><td>Invivogen</td><td>NC9141851</td><td>500x seyreltme</td></tr><tr><td>DAPI ile ProLong Gold Antifade Reaktifi</td><td>Hücre Sinyal Teknolojisi</td><td>8961S</td><td>Kullanıma hazır</td></tr><tr><td>Arındırılmış anti-fare CD16/32 Antikor</td><td>BioLegend</td><td>101301</td><td>100x seyreltme</td></tr><tr><td>RIPA buffer&nbsp;</td><td>Fisher Scientific</td><td>PI89900</td><td>Suda 10 kez seyreltme</td></tr><tr><td>Spektral akış sitometresi (Aurora 3)</td><td>Cytek Biosciences</td><td>Yok</td><td></td></tr><tr><td>Syntaxin 4 antikoru</td><td>Sinaptik Sistemler</td><td>110042</td><td>5000x seyrelme</td></tr><tr><td>Tecnai 20 (Geçirim Elektron Mikroskopu)</td><td>FEI</td><td>Yok</td><td></td></tr><tr><td>Tergeo-EM</td><td>PIE Scientific LLC</td><td>Yok</td><td></td></tr><tr><td>Vinculin antikoru</td><td>abcam</td><td>AB18508</td><td>5000x seyrelme</td></tr></tbody></table>

cd63-gfp raporlayıcı fare modeli kullanılarak adipositten türetilen ekstraselüler vezikül salgısının karakterizasyonu in vivo ve in vitro

Characterization of Adipocyte-Derived Extracellular Vesicle Secretion Using a CD63-GFP Reporter Mouse Model In Vivo and In Vitro

Konfokal mikroskopi ile APC alımıyla AdEV'lerin tespit edilmesi protokolünü kullanarak, AdipCD63-GFP ve Stopfl/fl/CD63-GFP farelerinin inguinal yağ dokularından APC'leri izole ettik ve çekirdeklerin görselleştirilmesi için DAPI ile boyadık. Konfokal mikroskopi ile incelendiğinde, Stopfl/fl/CD63-GFP farelerinden alınan APC'lerde tespit edilebilir GFP noktası bulunmamıştır (Şekil 1A). Buna karşılık, AdipCD63-GFP farelerinden izole edilen APC'ler, sitoplazma boyunca çok sa...

CD63-GFP Raporlayıcı Fare Modeli Kullanılarak Adipositten Türetilen Ekstraselüler Vezikül Salgısının Karakterizasyonu In Vivo ve In Vitro

Adipocytes are increasingly recognized as active endocrine cells that secrete substantial quantities of EVs enriched in proteins, lipids, and nucleic ...

characterization-adipocyte-derived-extracellular-vesicle-secretion

Research

JoVE Journal

Biology

Biyoloji

Endotelyal endotel / glycocalyx yüzey tabakası ideal intravital mikroskobu kullanılarak incelenmiştir. Intravital mikroskop teknik gibi akciğer gibi hareketli bir organ zordur. Biz eş zamanlı aydınlık ve floresan mikroskop serbestçe-hareket eden endotel yüzey tabakasının kalınlığını belirlemede nasıl kullanılabileceğini...

Fare Pulmoner Endotelyal Yüzey Katman in vivo ölçümü olarak

Bu protokol, adherent / mezenkimal meme epitel hücrelerinden ekstraselüler veziküller (EV) / ekzozomların arıtılması, nicelendirilmesi ve karakterize edilmesine yönelik yöntemleri ve meme bezi oluşturma yeteneğini luminal meme epitel hücrelerine aktarmak için kullanmayı açıklamaktadır. Gövde benzeri meme epitel hücrelerinden türetilmiş EVs / eksozomlar, bu hücre özelliğini EV'leri / eksosomları emen hücrelere aktarabilir.

Membran Bazal Epitelyal Hücreler ile Membran Luminal Hücreler Arasındaki Meme Bezi Oluşturma Yeteneğinin Ekstraselüler Vesiküller / Ekzozomlar Yoluyla Transferi

Yağ dokusu vasküler ataları gelen beyaz ve bej adiposit farklılaşma obezite metabolik iyileştirme için potansiyel taşımaktadır. CD34 + CD31 + insan yağı endotel hücre izolasyon ve bir sonraki vitro genişleme ve farklılaşma beyaz ve bej adiposit içine iletişim kurallarını açıklar. Birkaç aşağı akım uygulamaları ele alınmıştır.

Yalıtım, genişleme ve Adipojenik indüksiyon CD34 + CD31 + insan Omental ve deri altı yağ dokusu endotel hücrelerden

Bu protokol için hızlı izole edilmesi insan tam kan üzerinden ekstraselüler veziküller ve belirli işaretleri karakterizasyonu için tam iş akışı floresans tabanlı nanopartikül izleme analizi ile açıklanmaktadır. Sunulan sonuçlar tekrarlanabilirlik yüksek düzeyde göstermek ve hücre kültür supernatants için ayarlanabilir.

İnsan kanı nanopartikül izleme analizi ile tek hücre dışı veziküller floresans tabanlı hızlı karakterizasyonu

Biz adipoz doku metabolik fenotip katkıda matris ve adiposit lerin rollerinin diseksiyon sağlayan bir 3D insan ekstrasellüler matriks-adiposit in vitro kültür sistemi açıklar.

Bir İnsan 3D Ekstrasellüler Matrix-Adiposit Kültür Modeli Matrix-Hücre Metabolik Crosstalk Çalışma için

En yeni nesil EV karakterizasyon araçları, aynı anda birden fazla parametre arasında tek EV analizi yapabilir. Nano-akış sitometrisi, etiketleme yapmadan 45 nm'den büyük tüm biyolojik parçacıkları ölçer ve çeşitli floresan etiketleme teknikleriyle alt popülasyonların spesifik özelliklerini tanımlar.

Nano-Flow Sitometri ile Tek Hücre Dışı Vezikül Transmembran Protein Karakterizasyonu

Mevcut protokol, temsili EV'leri (eksozomlar ve mikro-parçacıklar) kültürlü insan MSC'lerinden izole etmek ve karakterize etmek için diferansiyel santrifüjlemeyi açıklamaktadır. Bu EV'lerin diğer uygulamaları da bu makalede açıklanmaktadır.

Kültürlenmiş İnsan Mezenkimal Kök Hücrelerinden Hücre Dışı Veziküllerin İzolasyonu, Karakterizasyonu ve Terapötik Uygulaması

Bu protokol, insan yağ kaynaklı mezenkimal kök hücrelerin (ADSC'ler) kültürlenmesinden ve süpernatantın toplanmasından ultrasantrifüjleme kullanılarak hücre dışı veziküllerin (EV'ler) çıkarılmasına kadar tüm prosedürleri açıklar.

İnsan Adipoz Kaynaklı Mezenkimal Kök Hücrelerden Hücre Dışı Veziküllerin Saflaştırılması ve Karakterizasyonu

Burada, insan deri altı yağ dokusundan (hSATMVEC'ler) mikrovasküler endotel hücrelerinin izole edilmesi, kültürlenmesi ve fenotiplendirilmesi için bir protokol tanımlanmıştır. Ek olarak, deneysel bir hSATMVEC-adiposit çapraz konuşma modelini açıklıyoruz.

İnsan Subkutan Yağ Dokusunda Adipoz Endotel Hücresi/Adiposit Çapraz Konuşmasının İncelenmesi

Ex vivo explant modeli kullanılarak, yağsız farelerden alınan visseral beyaz yağ dokusunun rostral kısmının küçük parçaları 7–10 gün boyunca kültürlenir; bu da yüksek yağlı diyet sırasında meydana gelen yağ dokusu iltihabının indüksiyonu ve incelenmesini mümkün kılar.

Yüksek Yağlı Diyet Benzeri Yağ Dokusu İltihaplanması için Deneyimli Fare Modeli

Keşfedin, CD63-GFP raporlayıcı fare modeli kullanarak adiposit kaynaklı ekstraselüler vezikül salgısını in vivo ve in vitro karakterize etme yöntemini. Bu protokol metabolik düzenleme ve hücre iletişimi araştırmaları için optimize edilmiş bir sistem sunar.

CD63-GFP Raporlayıcı Fare Modeli Kullanılarak Adipositten Türetilen Ekstraselüler Vezikül Salgısının Karakterizasyonu In Vivo ve In Vitro (Video) | JoVE Journal

CD63-GFP Raporlayıcı Fare Modeli Kullanılarak Adipositten Türetilen Ekstraselüler Vezikül Salgısının Karakterizasyonu In Vivo ve In Vitro

Daha fazla video keşfet