Name: Isolamento di cellule epiteliali primarie del tubulo prossimale umano e loro utilizzo nella creazione di un modello microfisiologico del tubulo prossimale renale (Video) | JoVE Journal
Uploaded: 2025-05-09T14:30:00.000+00:00
Duration: 7 min 6 s
Description: Scopri come isolare cellule epiteliali primarie del tubulo prossimale umano e utilizzarle in un sistema microfisiologico per valutare il trasporto di farmaci e la nefrotossicità. Questo metodo migliora la previsione delle risposte renali.

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>Bertram, J. F., Douglas-Denton, R. N., Diouf, B., Hughson, M. D., Hoy, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+nephron+number%3A+implications+for+health+and+disease%5BTitle%5D)+AND+%22Pediatr+Nephrol%22%5BJournal%5D)">Human nephron number: implications for health and disease.</a> Pediatr Nephrol. 26 (9), 1529-1533 (2011).</li>
<li>Zhang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Tubular+secretion+of+creatinine+and+kidney+function%3A+an+observational+study%5BTitle%5D)+AND+%22BMC+Nephrol%22%5BJournal%5D)">Tubular secretion of creatinine and kidney function: an observational study.</a> BMC Nephrol. 21, 108(2020).</li>
<li>Wang, K., Kestenbaum, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Proximal+tubular+secretory+clearance%5BTitle%5D)+AND+%22Clin+J+Am+Soc+Nephrol%22%5BJournal%5D)">Proximal tubular secretory clearance.</a> Clin J Am Soc Nephrol. 13 (8), 1291-1296 (2018).</li>
<li>Yin, J., Wang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Renal+drug+transporters+and+their+significance+in+drug-drug+interactions%5BTitle%5D)+AND+%22Acta+Pharm+Sin+B%22%5BJournal%5D)">Renal drug transporters and their significance in drug-drug interactions.</a> Acta Pharm Sin B. 6 (5), 363-373 (2016).</li>
<li>Connor, S., Roberts, R. A., Tong, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Drug-induced+kidney+injury%3A+challenges+and+opportunities%5BTitle%5D)+AND+%22Toxicol+Res+%28Camb%29%22%5BJournal%5D)">Drug-induced kidney injury: challenges and opportunities.</a> Toxicol Res (Camb). 13 (4), tfae119(2024).</li>
<li>Meijer, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+of+organic+anion+and+cation+transport+in+three+human+renal+proximal+tubular+epithelial+models%5BTitle%5D)+AND+%22Cells%22%5BJournal%5D)">Characterization of organic anion and cation transport in three human renal proximal tubular epithelial models.</a> Cells. 13 (12), 1008(2024).</li>
<li>Theocharis, A. D., Skandalis, S. S., Gialeli, C., Karamanos, N. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Extracellular+matrix+structure%5BTitle%5D)+AND+%22Adv+Drug+Deliv+Rev%22%5BJournal%5D)">Extracellular matrix structure.</a> Adv Drug Deliv Rev. 97, 4-27 (2016).</li>
<li>Miceli, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fluid+flow-induced+shear+stress+controls+the+metabolism+of+proximal+tubule+kidney+epithelial+cells+through+primary+cilium-dependent+lipophagy+and+mitochondria+biogenesis%5BTitle%5D)+AND+%22Autophagy%22%5BJournal%5D)">Fluid flow-induced shear stress controls the metabolism of proximal tubule kidney epithelial cells through primary cilium-dependent lipophagy and mitochondria biogenesis.</a> Autophagy. 16 (12), 2287-2288 (2020).</li>
<li>Tsang, Y. P., Hao, T., Mao, Q., Kelly, E. J., Unadkat, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dysregulation+of+the+mRNA+expression+of+human+renal+drug+transporters+by+proinflammatory+cytokines+in+primary+human+proximal+tubular+epithelial+cells%5BTitle%5D)+AND+%22Pharmaceutics%22%5BJournal%5D)">Dysregulation of the mRNA expression of human renal drug transporters by proinflammatory cytokines in primary human proximal tubular epithelial cells.</a> Pharmaceutics. 16 (2), 285(2024).</li>
<li>Bajaj, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Freshly+isolated+primary+human+proximal+tubule+cells+as+an+in+vitro+model+for+the+detection+of+renal+tubular+toxicity%5BTitle%5D)+AND+%22Toxicology%22%5BJournal%5D)">Freshly isolated primary human proximal tubule cells as an in vitro model for the detection of renal tubular toxicity.</a> Toxicology. 442, 152535(2020).</li>
<li>Brown, C. D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterisation+of+human+tubular+cell+monolayers+as+a+model+of+proximal+tubular+xenobiotic+handling%5BTitle%5D)+AND+%22Toxicol+Appl+Pharmacol%22%5BJournal%5D)">Characterisation of human tubular cell monolayers as a model of proximal tubular xenobiotic handling.</a> Toxicol Appl Pharmacol. 233 (3), 428-438 (2008).</li>
<li>Becker, G. J., Hewitson, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Animal+models+of+chronic+kidney+disease%3A+useful+but+not+perfect%5BTitle%5D)+AND+%22Nephrol+Dial+Transplant%22%5BJournal%5D)">Animal models of chronic kidney disease: useful but not perfect.</a> Nephrol Dial Transplant. 28 (10), 2432-2438 (2013).</li>
<li>Morse, B. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pharmacokinetics+of+organic+cation+transporter+1+%28OCT1%29+substrates+in+Oct1%2F2+knockout+mice+and+species+difference+in+hepatic+OCT1-mediated+uptake%5BTitle%5D)+AND+%22Drug+Metab+Dispos%22%5BJournal%5D)">Pharmacokinetics of organic cation transporter 1 (OCT1) substrates in Oct1/2 knockout mice and species difference in hepatic OCT1-mediated uptake.</a> Drug Metab Dispos. 48 (2), 93-105 (2020).</li>
<li>Samodelov, S. L., Kullak-Ublick, G. A., Gai, Z., Visentin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Organic+cation+transporters+in+human+physiology%2C+pharmacology%2C+and+toxicology%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Organic cation transporters in human physiology, pharmacology, and toxicology.</a> Int J Mol Sci. 21 (21), 7890(2020).</li>
<li>Wikswo, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+relevance+and+potential+roles+of+microphysiological+systems+in+biology+and+medicine%5BTitle%5D)+AND+%22Exp+Biol+Med%22%5BJournal%5D)">The relevance and potential roles of microphysiological systems in biology and medicine.</a> Exp Biol Med. 239 (9), 1061-1072 (2014).</li>
<li>Weber, E. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Development+of+a+microphysiological+model+of+human+kidney+proximal+tubule+function%5BTitle%5D)+AND+%22Kidney+Int%22%5BJournal%5D)">Development of a microphysiological model of human kidney proximal tubule function.</a> Kidney Int. 90 (3), 627-637 (2016).</li>
<li>Chapron, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+improved+vascularized%2C+dual-channel+microphysiological+system+facilitates+modeling+of+proximal+tubular+solute+secretion%5BTitle%5D)+AND+%22ACS+Pharmacol+Transl+Sci%22%5BJournal%5D)">An improved vascularized, dual-channel microphysiological system facilitates modeling of proximal tubular solute secretion.</a> ACS Pharmacol Transl Sci. 3 (3), 496-508 (2020).</li>
<li>Adler, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+quantitative+approach+to+screen+for+nephrotoxic+compounds+in+vitro%5BTitle%5D)+AND+%22J+Am+Soc+Nephrol%22%5BJournal%5D)">A quantitative approach to screen for nephrotoxic compounds in vitro.</a> J Am Soc Nephrol. 27 (4), 1015-1028 (2016).</li>
<li>Chang, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+liver-kidney+model+elucidates+the+mechanisms+of+aristolochic+acid+nephrotoxicity%5BTitle%5D)+AND+%22JCI+Insight%22%5BJournal%5D)">Human liver-kidney model elucidates the mechanisms of aristolochic acid nephrotoxicity.</a> JCI Insight. 2 (22), e95978(2017).</li>
<li>Weber, E. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+kidney+on+a+chip+assessment+of+polymyxin+antibiotic+nephrotoxicity%5BTitle%5D)+AND+%22JCI+Insight%22%5BJournal%5D)">Human kidney on a chip assessment of polymyxin antibiotic nephrotoxicity.</a> JCI Insight. 3 (24), e123673(2018).</li>
<li>Imaoka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bridging+the+gap+between+in+silico+and+in+vivo+by+modeling+opioid+disposition+in+a+kidney+proximal+tubule+microphysiological+system%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Bridging the gap between in silico and in vivo by modeling opioid disposition in a kidney proximal tubule microphysiological system.</a> Sci Rep. 11 (1), 21356(2021).</li>
<li>Van Ness, K. P., Chang, S., Weber, E. J., Zumpano, D., Eaton, D. L., Kelly, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microphysiological+systems+to+assess+nonclinical+toxicity%5BTitle%5D)+AND+%22Curr+Protoc+Toxicol%22%5BJournal%5D)">Microphysiological systems to assess nonclinical toxicity.</a> Curr Protoc Toxicol. 73, 14.18.1-14.18.28 (2017).</li>
<li>Lidberg, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Serum+protein+exposure+activates+a+core+regulatory+program+driving+human+proximal+tubule+injury%5BTitle%5D)+AND+%22J+Am+Soc+Nephrol%22%5BJournal%5D)">Serum protein exposure activates a core regulatory program driving human proximal tubule injury.</a> J Am Soc Nephrol. 33 (5), 949-965 (2022).</li>
<li>Sakolish, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Technology+transfer+of+the+microphysiological+systems%3A+a+case+study+of+the+human+proximal+tubule+tissue+chip%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Technology transfer of the microphysiological systems: a case study of the human proximal tubule tissue chip.</a> Sci Rep. 8 (1), 14882(2018).</li>
<li>Hart, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Identification+of+prognostic+biomarkers+for+antibiotic+associated+nephrotoxicity+in+cystic+fibrosis%5BTitle%5D)+AND+%22J+Cyst+Fibros%22%5BJournal%5D)">Identification of prognostic biomarkers for antibiotic associated nephrotoxicity in cystic fibrosis.</a> J Cyst Fibros. 23 (2), 293-299 (2024).</li>
<li>Lidberg, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antisense+oligonucleotide+development+for+the+selective+modulation+of+CYP3A5+in+renal+disease%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Antisense oligonucleotide development for the selective modulation of CYP3A5 in renal disease.</a> Sci Rep. 11 (1), 4722(2021).</li>
<li>Imaoka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microphysiological+system+modeling+of+ochratoxin+A-associated+nephrotoxicity%5BTitle%5D)+AND+%22Toxicology%22%5BJournal%5D)">Microphysiological system modeling of ochratoxin A-associated nephrotoxicity.</a> Toxicology. 444, 152582(2020).</li>
<li>Chapron, B. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reevaluating+the+role+of+megalin+in+renal+vitamin+D+homeostasis+using+a+human+cellderived+microphysiological+system%5BTitle%5D)+AND+%22ALTEX%22%5BJournal%5D)">Reevaluating the role of megalin in renal vitamin D homeostasis using a human cellderived microphysiological system.</a> ALTEX. 35 (4), 504-515 (2018).</li>
<li>Adashi, E. Y., O'Mahony, D. P., Cohen, I. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+FDA+modernization+act+2.0%3A+drug+testing+in+animals+is+rendered+optional%5BTitle%5D)+AND+%22Am+J+Med%22%5BJournal%5D)">The FDA modernization act 2.0: drug testing in animals is rendered optional.</a> Am J Med. 136 (9), 853-854 (2023).</li>
<li>Qi, W., Johnson, D. W., Vesey, D. A., Pollock, C. A., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation%2C+propagation+and+characterization+of+primary+tubule+cell+culture+from+human+kidney+%28methods+in+renal+research%29%5BTitle%5D)+AND+%22Nephrology%22%5BJournal%5D)">Isolation, propagation and characterization of primary tubule cell culture from human kidney (methods in renal research).</a> Nephrology. 12 (2), 155-159 (2007).</li>
<li>Mihevc, M., Petreski, T., Maver, U., Bevc, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Renal+proximal+tubular+epithelial+cells%3A+review+of+isolation%2C+characterization%2C+and+culturing+techniques%5BTitle%5D)+AND+%22Mol+Biol+Rep%22%5BJournal%5D)">Renal proximal tubular epithelial cells: review of isolation, characterization, and culturing techniques.</a> Mol Biol Rep. 47 (12), 9865-9882 (2020).</li>
<li>Shaver, C. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cellfree+hemoglobin+augments+acute+kidney+injury+during+experimental+sepsis%5BTitle%5D)+AND+%22Am+J+Physiol+Renal+Physiol%22%5BJournal%5D)">Cellfree hemoglobin augments acute kidney injury during experimental sepsis.</a> Am J Physiol Renal Physiol. 317 (4), F922-F929 (2019).</li>
<li>Sbarbati, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Separation+of+human+endothelial+cells+from+fibroblasts+by+centrifugation+in+Percoll+gradients%5BTitle%5D)+AND+%22Biosci+Rep%22%5BJournal%5D)">Separation of human endothelial cells from fibroblasts by centrifugation in Percoll gradients.</a> Biosci Rep. 5 (6), 469-472 (1985).</li>
<li>Horn, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+of+human+mesenchymal+stromal+cells+is+more+efficient+by+red+blood+cell+lysis%5BTitle%5D)+AND+%22Cytotherapy%22%5BJournal%5D)">Isolation of human mesenchymal stromal cells is more efficient by red blood cell lysis.</a> Cytotherapy. 10 (7), 676-685 (2008).</li>
</ol>

<table><tbody><tr><td>1 mL di siringhe Luer-Lok Tip</td><td>Becton, Dickinson</td><td>309628</td><td></td></tr><tr><td>Tubi microcentrifughe da 1,5 mL</td><td>Prodotti Scientifici CELLTREAT</td><td>229442</td><td></td></tr><tr><td>Tubo sterile di polipropilene conico da 15 mL</td><td>Fisher Scientific</td><td>14-959-53A</td><td></td></tr><tr><td>Dispositivo microfisiologico 3D</td><td>Nortis</td><td>TCC-001</td><td></td></tr><tr><td>Siringhe Luer-Lok Tip da 5 mL</td><td>Becton, Dickinson</td><td>309646</td><td></td></tr><tr><td>Tubo conico sterile in polipropilene da 50 mL</td><td>Fisher Scientific</td><td>12-565-271</td><td></td></tr><tr><td>Antibiotico-Antimicotico (100 volte)</td><td>ThermoFisher Scientific</td><td>15240062</td><td></td></tr><tr><td>Collagenase, tipo IV, polvere</td><td>ThermoFisher Scientific</td><td>17104019</td><td></td></tr><tr><td>D-(+)-Glucosio</td><td>MilliporeSigma</td><td>G8270</td><td></td></tr><tr><td>Inibitore della tripsina definito</td><td>ThermoFisher Scientific</td><td>R007100</td><td></td></tr><tr><td>Dimetilsolfosde, bioreagente, Thermo Scientific Chemicals</td><td>ThermoFisher Scientific</td><td>J66650-AK</td><td></td></tr><tr><td>Dispase II, polvere</td><td>ThermoFisher Scientific</td><td>17105041</td><td></td></tr><tr><td>Dulbecco' s MEM (DMEM)/F-12 senza glucosio</td><td>Scienze della Vita Biologiche degli Stati Uniti</td><td>D9807-02</td><td></td></tr><tr><td>Soluzione salina tamponata con fosfato di Dulbecco (DPBS), calcio, magnesio</td><td>ThermoFisher Scientific</td><td>14040117</td><td></td></tr><tr><td>Siero Fetale Bovino (FBS), Premium</td><td>ThermoFisher Scientific</td><td>A5670701</td><td></td></tr><tr><td>Kit Finnpipette F2 Good Laboratory Pipetting (GLP)</td><td>ThermoFisher Scientific</td><td>05-719-511</td><td>Micropipette</td></tr><tr><td>Punte per pipette Fisherbrand SureOne Micropoint, Universal Fit, Non Filtrate</td><td>Fisher Scientific</td><td>02707407, 02707410, 02707438</td><td></td></tr><tr><td>Rene umano intero fresco</td><td>Novabiosi</td><td>https://www.novabiosis.com/research-organ-allocation-services/</td><td>I reni umani sono stati ottenuti tramite Novabiosis, un OPO approvato da UNS</td></tr><tr><td>Incubatore umidificato (37 &deg; C e 5% CO2)</td><td>Fisher Scientific</td><td>51033556</td><td></td></tr><tr><td>Idrocortisone</td><td>MilliporeSigma</td><td>H0888</td><td></td></tr><tr><td>Shaker orbitale incubante</td><td>Avantor</td><td>12620-946</td><td></td></tr><tr><td>Insulina-Transferrina-Selenio (100X) (ITS -G)</td><td>ThermoFisher Scientific</td><td>41400045</td><td></td></tr><tr><td>Fiale criogeniche a filo interno</td><td>Corning</td><td>430487</td><td></td></tr><tr><td>Kimtech Scienza Kimwipes</td><td>Kimberly-Clark Professionista</td><td>34120</td><td></td></tr><tr><td>Luer Stubs (ago smussato), 22 G x 0,5 pollici</td><td>Instech Laboratories, Inc.</td><td>LS22</td><td></td></tr><tr><td>Medium 199, Sali di Earle (10 volte)</td><td>ThermoFisher Scientific</td><td>21180021</td><td></td></tr><tr><td>Megafuge 8 Piccola centrifuga da banco</td><td>ThermoFisher Scientific</td><td>75007210</td><td></td></tr><tr><td>Accoppiatore metallico (smussato)</td><td>Instech Laboratories, Inc.</td><td>SC 20/15</td><td></td></tr><tr><td>Signor Ghiaccio Ghiacciante</td><td>ThermoFisher Scientific</td><td>5100-0036</td><td></td></tr><tr><td>Microscopio Nikon Eclipse Ti-S</td><td>Strumenti Nikon</td><td>https://www.thelabworldgroup.com/product/nikon-eclipse-ti-fluorescent-microscope/</td><td>Modello originale dismesso&nbsp;</td></tr><tr><td>Fiaschette Non trattate Nunc, T-25</td><td>ThermoFisher Scientific</td><td>169900</td><td></td></tr><tr><td>Nunc Fiaschette non trattate, T-75</td><td>ThermoFisher Scientific</td><td>156800</td><td></td></tr><tr><td>Pipetta, 10 mL, Graduata, 1/10 mL, Sterile</td><td>Greiner Bio-One</td><td>607180</td><td></td></tr><tr><td>Tubo in silicio, tubo C-flex, (ID: 0,020", OD: 0,083")</td><td>Cole-Parmer</td><td>06422-00</td><td></td></tr><tr><td>Lametta a filo singolo (sterile)</td><td>Biosigillo</td><td>KI-205/50</td><td></td></tr><tr><td>Bicarbonato di sodio</td><td>MilliporeSigma</td><td>S6297</td><td></td></tr><tr><td>Idrossido di sodio</td><td>MilliporeSigma</td><td>S8045</td><td></td></tr><tr><td>Unità di filtraggio sterili usa e getta con membrane PES</td><td>ThermoFisher Scientific</td><td>567-0020</td><td></td></tr><tr><td>Piatti trattati con coltura tissutale, 150 mm&nbsp; x 20 mm ventilato</td><td>Genesee Scientific</td><td>25-203</td><td></td></tr><tr><td>Tripsina-EDTA (0,05%), fenolo rosso</td><td>ThermoFisher Scientific</td><td>25300054</td><td></td></tr><tr><td>Collagene di tipo I, coda di ratto</td><td>Corning</td><td>354236</td><td></td></tr><tr><td>Collagene di tipo IV, topo</td><td>Corning</td><td>354233</td><td></td></tr></tbody></table>

isolamento di cellule epiteliali primarie del tubulo prossimale umano e loro utilizzo nella creazione di un modello microfisiologico del tubulo prossimale renale

Isolation of Primary Human Proximal Tubule Epithelial Cells and Their Use in Creating a Microphysiological Model of the Renal Proximal Tubule

Isolamento di cellule epiteliali primarie del tubulo prossimale umano e loro utilizzo nella creazione di un modello microfisiologico del tubulo prossimale renale

Kidney disease affects over 850 million people globally, including 37 million Americans. Risk factors for&#160;chronic kidney disease include ...

isolation-primary-human-proximal-tubule-epithelial-cells-their-use

Research

Ricerca

JoVE Journal

Medicine

Medicina

In questo video, dimostriamo i passaggi per isolare e purificare le cellule epiteliali tubulari renali per un'ampia gamma di studi di biologia molecolare e colture cellulari.

Isolamento di cellule renali mure: una tecnica per isolare e generare colture primarie di cellule epiteliali tubulari renali

Canali ionici espressi in renale tubulare svolgono un ruolo significativo nella patologia della malattia del rene policistico. Qui si descrive protocolli sperimentali utilizzati per eseguire patch-clamp analisi e di livello di calcio intracellulare misure in cistico appena isolato da reni di roditori.

Implementazione Patch Clamp e vivo microscopia a fluorescenza per il monitoraggio delle proprietà funzionali di recente isolato PKD Epithelium

Questo protocollo descrive la decellularizzazione dei reni di ratto Sprague Dawley mediante perfusione anterograda di detergenti attraverso il sistema vascolare, producendo matrici extracellulari renali acellulari che fungono da modelli per il ripopolamento con cellule epiteliali renali umane. La ricellularizzazione e l'uso del saggio di perfusione della resazurina per monitorare la crescita vengono eseguiti all'interno di bioreattori di perfusione appositamente progettati.

Ripopolamento cellule epiteliali e preparazione di roditori matrice extracellulare Ponteggi per lo sviluppo del tessuto renale

Qui presentiamo un metodo di isolamento e coltura di romanzo grado clinico per rene perivascolare di cellule stromali (kPSCs) basato sulla perfusione d'organo intero con gli enzimi digestivi e arricchimento NG2-cella. Con questo metodo, è possibile acquisire il numero di celle sufficiente per una terapia cellulare.

Un metodo di isolamento di romanzo Clinical Grade per rene umano perivascolari cellule stromali

Tessutale renale costrutti forniscono una soluzione per la carenza di organi e deleteri effetti della dialisi. Qui, descriviamo un protocollo a micro sezionare murini reni per isolamento di segmenti cortico-midollare. Questi segmenti sono impiantati nei costrutti senza impalcatura cellulare, formando organoids renale.

Microdissection di segmenti di tessuto renale primario e incorporazione con tecnologia romanzo costrutto senza impalcatura

Questo protocollo fornisce un approccio conveniente per isolare e caratterizzare del mouse primario cellule tubolari renali che possono successivamente essere sub-coltivate per valutare funzioni biologiche renale ex vivo, tra cui bioenergetiche mitocondriali.

Isolamento, caratterizzazione e Throughput elevato flusso extracellulare analisi di cellule epiteliali tubolari renali primari del Mouse

Le colture tubuloidi renali umane rappresentano un prezioso modello in vitro per studiare la fisiologia e la malattia renale. I tubuloidi possono essere stabiliti dal tessuto renale (sano e malato) e dall'urina, quest'ultima rappresenta una fonte facilmente ottenibile e meno invasiva di materiale di ricerca.

Generazione di tubuloidi renali umani da tessuti e urine

Qui, presentiamo protocolli che descrivono in dettaglio le istruzioni per l'implementazione di colture cellulari 3D utilizzando matrici di collagene e collagene-agarosio in un sistema microfisiologico. Questi protocolli supportano la co-coltura del tubulo prossimale renale e degli sferoidi del carcinoma a cellule renali, simulando condizioni in vivo e consentendo un'indagine avanzata sulle interazioni tra le cellule tumorali del rene.

Co-coltura microfluidica dell'epitelio renale sano e tumorale per modellare la progressione del cancro del rene

Qui, descriviamo i protocolli per ottenere colture primarie di cellule epiteliali tubulari renali umane (HRTEC) dalla corteccia renale per sviluppare monostrati e colture tridimensionali su una matrice extracellulare (3D-HRTEC).

Sviluppo di colture primarie di cellule epiteliali tubulari renali umane in monostrati e condizioni tridimensionali

Qui descriviamo un protocollo in vitro per modellare danni da ipossia/reossigenazione nelle cellule epiteliali tubulari prossimali altamente metaboliche. Questo modello è progettato per indurre danni indotti dallo stress metabolico, che possono essere misurati attraverso l'espressione di marcatori di danno prossimale dei tubuli e la valutazione della funzione respiratoria mitocondriale.

Modellizzazione dell'ipossia/lesione da reossigenazione nelle cellule epiteliali tubolari prossimali

Scopri come isolare cellule epiteliali primarie del tubulo prossimale umano e utilizzarle in un sistema microfisiologico per valutare il trasporto di farmaci e la nefrotossicità. Questo metodo migliora la previsione delle risposte renali.

Isolamento di cellule epiteliali primarie del tubulo prossimale umano e loro utilizzo nella creazione di un modello microfisiologico del tubulo prossimale renale (Video) | JoVE Journal

Isolamento di cellule epiteliali primarie del tubulo prossimale umano e loro utilizzo nella creazione di un modello microfisiologico del tubulo prossimale renale

Esplora altri video