Name: transfer of mammary gland-forming ability between mammary basal epithelial cells and mammary luminal cells via extracellular vesicles/exosomes
Uploaded: 2017-06-03T14:00:00
Description: Watch this Scientific Journal Video about Transfer of Mammary Gland-forming Ability Between Mammary Basal Epithelial Cells and Mammary Luminal Cells via Extracellular Vesicles/Exosomes at JoVE.com

Questo lavoro è stato sostenuto da sovvenzioni degli Istituti Nazionali di Ricerca Sanitaria (05A1-CSPP16-014, HJL) e dal Ministero della Scienza e della Tecnologia (MOST 103-2320-B-400-015-MY3, HJL).

Tutte le ricerche relative agli animali sono conformi ai protocolli approvati dal Comitato istituzionale per la cura degli animali. 1. Isolamento e validazione di vescica extracellulare / esosomi <ol><li> Le cellule basali dell&#39;epitelio mammario della cultura, NAMECs 4 , con mezzo fresco e privo di siero costituito da 500 mL di MCDB 170, pH 7.4 + 500 mL di DMEM / F12 con bicarbonato di sodio (0,2438%); EGF (5 ng / mL); Idrocortisone (0,5 μg / mL); Insulina (5 ?...

<ol>
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C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PGE2+/EP4+Signaling+Controls+the+Transfer+of+the+Mammary+Stem+Cell+State+by+Lipid+Rafts+in+Extracellular+Vesicles.">PGE2 /EP4 Signaling Controls the Transfer of the Mammary Stem Cell State by Lipid Rafts in Extracellular Vesicles.</a> Stem Cells. , (2016).</li><li>Th&#233;ry, C., Amigorena, S., Raposo, G., Clayton, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+exosomes+from+cell+culture+supernatants+and+biological+fluids.">Isolation and characterization of exosomes from cell culture supernatants and biological fluids.</a> Curr Protoc Cell Biol. , (2006).</li><li>Gy&#246;rgy, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+vesicles,+current+state-of-the-art:+emerging+role+of+extracellular+vesicles.">Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles.</a> Cell Mol Life Sci. 68 (16), 2667-2688 (2011).</li><li>Olver, C., Vidal, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteomic+analysis+of+secreted+exosomes.">Proteomic analysis of secreted exosomes.</a> Subcell Biochem. 43, 99-131 (2007).</li><li>Shackleton, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+a+functional+mammary+gland+from+a+single+stem+cell.">Generation of a functional mammary gland from a single stem cell.</a> Nature. 439 (7072), 84-88 (2006).</li><li>Prater, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mammary+stem+cells+have+myoepithelial+cell+properties.">Mammary stem cells have myoepithelial cell properties.</a> Nat Cell Biol. 16 (10), 942-950 (2014).</li><li>Stingl, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purification+and+unique+properties+of+mammary+epithelial+stem+cells.">Purification and unique properties of mammary epithelial stem cells.</a> Nature. 439 (7079), 993-997 (2006).</li><li>Gardiner, C., Ferreira, Y. J., Dragovic, R. A., Redman, C. W., Sargent, I. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+vesicle+sizing+and+enumeration+by+nanoparticle+tracking+analysis.">Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis.</a> J Extracell Vesicles. 2, (2013).</li><li>Shapiro, A. L., Vi&#241;uela, E., Maizel, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+weight+estimation+of+polypeptide+chains+by+electrophoresis+in+SDS-polyacrylamide+gels.">Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels.</a> Biochem Biophys Res Commun. 28 (5), 815-820 (1967).</li><li>Riches, A., Campbell, E., Borger, E., Powis, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+exosome+release+from+mammary+epithelial+and+breast+cancer+cells+-+a+new+regulatory+pathway.">Regulation of exosome release from mammary epithelial and breast cancer cells - a new regulatory pathway.</a> Eur J Cancer. 50 (5), 1025-1034 (2014).</li><li>Witwer, K. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardization+of+sample+collection,+isolation+and+analysis+methods+in+extracellular+vesicle+research.">Standardization of sample collection, isolation and analysis methods in extracellular vesicle research.</a> J Extracell Vesicles. 2, (2013).</li><li>van der Vlist, E. J., Nolte-'t Hoen, E. N., Stoorvogel, W., Arkesteijn, G. J., Wauben, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescent+labeling+of+nano-sized+vesicles+released+by+cells+and+subsequent+quantitative+and+qualitative+analysis+by+high-resolution+flow+cytometry.">Fluorescent labeling of nano-sized vesicles released by cells and subsequent quantitative and qualitative analysis by high-resolution flow cytometry.</a> Nat Protoc. 7 (7), 1311-1326 (2012).</li><li>Kowal, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteomic+comparison+defines+novel+markers+to+characterize+heterogeneous+populations+of+extracellular+vesicle+subtypes.">Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes.</a> Proc Natl Acad Sci U S A. 113 (8), E968-E977 (2016).</li><li>Li, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ADVANCED+IMAGING.+Extended-resolution+structured+illumination+imaging+of+endocytic+and+cytoskeletal+dynamics.">ADVANCED IMAGING. Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics.</a> Science. 349 (6251), (2015).</li><li>Outzen, H. C., Custer, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+of+human+normal+and+neoplastic+mammary+tissues+in+the+cleared+mammary+fat+pad+of+the+nude+mouse.">Growth of human normal and neoplastic mammary tissues in the cleared mammary fat pad of the nude mouse.</a> J Natl Cancer Inst. 55 (6), 1461-1466 (1975).</li><li>Sheffield, L. G., Welsch, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+human+breast+epithelia+to+mammary-gland-free+fat-pads+of+athymic+nude+mice:+influence+of+mammotrophic+hormones+on+growth+of+breast+epithelia.">Transplantation of human breast epithelia to mammary-gland-free fat-pads of athymic nude mice: influence of mammotrophic hormones on growth of breast epithelia.</a> Int J Cancer. 41 (5), 713-719 (1988).</li></ol>

Gli esosomi spesso trasportano caratteristiche delle cellule che li hanno liberati e la quantità di esosomi rilasciati può essere indotta da stimoli 4 . Il mezzo di coltura delle cellule può essere raccolto e sottoposto ad ultracentrifugazione differenziale per la raccolta di EV / esosomi ( Figura 1 ). Attualmente non esiste un accordo generale su un metodo ideale per isolare EV / esosomi. Il metodo ottimale utilizzato qui è stato determinato dall&#39;applicazion...

Gli esosomi possono mediare la comunicazione cellulare trasferendo membrane e proteine ​​citosoliche, lipidi e RNA tra le cellule 1 . La comunicazione esosomiale è stata dimostrata coinvolta in molti processi fisiologici e patologici ( es. Presentazione di antigene, sviluppo della tolleranza 2 e progressione tumorale 3 ). Gli esosomi spesso hanno contenuti simili a quelli delle cellule di origine che li rilasciano. Così, gli esosomi possono trasportare proprietà cellulari specifiche dalle cellule sorgenti e trasferire queste proprietà alle cellule che ingeriscono 4 . Gli esosomi sono vescicole a doppio strato da 50 a 150 nm e presentano marcatori specifici ( es. CD9, CD81, CD63, HSP70, Alix e TSG101). Così, gli esosomi devono essere caratterizzati da vari metodi per diversi aspetti. La microscopia elettronica di trasmissione può essere usata per visualizzare le vescicole a membranaQuali esosomi 4 , 5 . L&#39;analisi di nanoparticle tracking (NTA) e l&#39;analisi della diffusione della luce dinamica (DLS) vengono utilizzati per misurare la dimensione e il numero di esosomi purificati 4 . Il contenuto di membrana lipidica degli esosomi può essere verificato mediante gradiente di densità. Marcatori esosomali, come CD9, CD81, CD63, HSP70, Alix e TSG101 6,7, possono essere misurati mediante blottatura Western. Le cellule basali del mammario hanno la capacità di generare ghiandole mammarie quando vengono impiantati in pastiglie grasse, mentre le cellule luminali non possono essere 8 , 9 , 10 . Così, le cellule basali del mammario sono anche definite come unità ripopolanti mammarie. Utilizzando il modello delle cellule mammarie basali e luminali, è possibile esaminare la capacità di EVs / esosomi di trasferire le caratteristiche delle cellule tra diverse popolazioni cellulari. Questo lavoroDimostra il metodo di trasferire la capacità di formazione delle ghiandole dalle cellule epiteliali basali del mammario alle cellule epiteliali luminescenti mammarie usando EVs / esosomi derivanti da cellule epiteliali basali mammarie. Le cellule epiteliali mammarie luminescenti hanno acquisito proprietà cellulari basali dopo l&#39;ingestione di EV / esosomi secretati dalle cellule basali e quindi possono formare ghiandole mammarie 4 .

<table>
	<tbody>
		<tr>
			<td>MCDB 170&#160;</td>
			<td>USBiological</td>
			<td>M2162</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F12</td>
			<td>Thermo</td>
			<td>1250062</td>
			<td></td>
		</tr>
		<tr>
			<td>Optima L-100K ultracentrifuge</td>
			<td>Beckman</td>
			<td>393253</td>
			<td></td>
		</tr>
		<tr>
			<td>SW28 Ti Rotor</td>
			<td>Beckman</td>
			<td>342204</td>
			<td></td>
		</tr>
		<tr>
			<td>SW41 Rotor</td>
			<td>Beckman</td>
			<td>331306</td>
			<td></td>
		</tr>
		<tr>
			<td>NANOSIGHT LM10</td>
			<td>Malvern</td>
			<td>NANOSIGHT LM10</td>
			<td>for nanoparticle tracking analysis (NTA)</td>
		</tr>
		<tr>
			<td>Optiprep&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>D1556</td>
			<td>60% (w/v) solution of iodixanol in water (sterile).</td>
		</tr>
		<tr>
			<td>CD81 antibody</td>
			<td>GeneTex</td>
			<td>GTX101766</td>
			<td>1:1000 in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween 20 at 4&#176;C, overnight&#160;</td>
		</tr>
		<tr>
			<td>CD9 antibody</td>
			<td>GeneTex</td>
			<td>GTX100912</td>
			<td>1:1000 in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween 20 at 4&#176;C, overnight&#160;</td>
		</tr>
		<tr>
			<td>CD63 antibody</td>
			<td>Abcam</td>
			<td>Ab59479</td>
			<td>1:1000 in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween 20 at 4&#176;C, overnight&#160;</td>
		</tr>
		<tr>
			<td>TSG101 antibody</td>
			<td>GeneTex</td>
			<td>GTX118736</td>
			<td>1:1000 in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween 20 at 4&#176;C, overnight&#160;</td>
		</tr>
		<tr>
			<td>GAPDH</td>
			<td>GeneTex</td>
			<td>GTX100118</td>
			<td>1:6000 in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween 20 at 4&#176;C, overnight&#160;</td>
		</tr>
		<tr>
			<td>CFSE (carboxyfluorescein succinimidyl diacetate ester)</td>
			<td>Thermo</td>
			<td>V12883</td>
			<td></td>
		</tr>
		<tr>
			<td>FACSCalibur</td>
			<td>BD Biosciences</td>
			<td></td>
			<td>fluorescence cell analyzer</td>
		</tr>
		<tr>
			<td>collagenase Type IV&#160;</td>
			<td>Thermo</td>
			<td>17104019</td>
			<td></td>
		</tr>
		<tr>
			<td>trypsin</td>
			<td>Thermo</td>
			<td>27250018</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;ITS</td>
			<td>Sigma-Aldrich</td>
			<td>I3146</td>
			<td>a mixture of recombinant human insulin, human transferrin, and sodium selenite</td>
		</tr>
		<tr>
			<td>accutase</td>
			<td>ebioscience</td>
			<td>00-4555-56</td>
			<td>a natural enzyme mixture with proteolytic and collagenolytic enzyme activity</td>
		</tr>
		<tr>
			<td>dispase&#160;</td>
			<td>STEMCELL</td>
			<td>7913</td>
			<td>5 mg/ml = 5 U/ml</td>
		</tr>
		<tr>
			<td>anti-CD49f antibody</td>
			<td>Biolegend</td>
			<td>313611</td>
			<td>1:50</td>
		</tr>
		<tr>
			<td>anti-EpCAM antibody</td>
			<td>Biolegend</td>
			<td>118213</td>
			<td>1:200</td>
		</tr>
		<tr>
			<td>FACSAria</td>
			<td>BD Biosciences</td>
			<td></td>
			<td>cell sorter</td>
		</tr>
		<tr>
			<td>carmine alum</td>
			<td>Sigma-Aldrich</td>
			<td>C1022</td>
			<td></td>
		</tr>
		<tr>
			<td>human mammary epithelial cells (HMLE cells, NAMECs)</td>
			<td></td>
			<td></td>
			<td>gifts from Dr. Robert Weinberg</td>
		</tr>
		<tr>
			<td>permount</td>
			<td>Thermo Fisher Scientific&#160;</td>
			<td>SP15-500</td>
			<td></td>
		</tr>
		<tr>
			<td>sodium bicarbonate</td>
			<td>Zymeset&#160;</td>
			<td>BSB101</td>
			<td></td>
		</tr>
		<tr>
			<td>EGF</td>
			<td>Peprotech</td>
			<td>AF-100-015</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrocoritisone</td>
			<td>Sigma-Aldrich</td>
			<td>SI-H0888</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>SI-I9278</td>
			<td></td>
		</tr>
		<tr>
			<td>BPE (bovine pituitary extract)</td>
			<td>Hammod Cell Tech&#160;</td>
			<td>1078-NZ</td>
			<td></td>
		</tr>
		<tr>
			<td>GW627368X&#160;</td>
			<td>Cayman</td>
			<td>10009162</td>
			<td></td>
		</tr>
		<tr>
			<td>15-cm culture dish</td>
			<td>Falcon&#160;</td>
			<td>353025</td>
			<td></td>
		</tr>
		<tr>
			<td>table-top centrifuge</td>
			<td>Eppendrof&#160;</td>
			<td>Centrifuge 3415R</td>
			<td></td>
		</tr>
		<tr>
			<td>ultracentrifuge tube</td>
			<td>Beckman</td>
			<td>344058</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS (Phosphate-buffered saline)&#160;</td>
			<td>Corning</td>
			<td>46-013-CM</td>
			<td></td>
		</tr>
		<tr>
			<td>BCA Protein Assay</td>
			<td>Thermo Fisher Scientific&#160;</td>
			<td>23228</td>
			<td></td>
		</tr>
		<tr>
			<td>Transmission Electron Microscopy</td>
			<td>Hitachi</td>
			<td>HT7700</td>
			<td></td>
		</tr>
		<tr>
			<td>gelatin&#160;</td>
			<td>STEMCELL</td>
			<td>7903</td>
			<td></td>
		</tr>
		<tr>
			<td>10-cm culture dish</td>
			<td>Falcon&#160;</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well culture dish</td>
			<td>Corning</td>
			<td>3516</td>
			<td></td>
		</tr>
		<tr>
			<td>female C57BL/6 mice</td>
			<td>NLAC (National Laboratory Animal Center</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>FBS (Fetal Bovine Serum)</td>
			<td>BioWest&#160;</td>
			<td>S01520</td>
			<td></td>
		</tr>
		<tr>
			<td>gentamycin</td>
			<td>Thermo Fisher Scientific&#160;</td>
			<td>15710072</td>
			<td></td>
		</tr>
		<tr>
			<td>Pen/Strep</td>
			<td>Corning</td>
			<td>30-002-Cl</td>
			<td></td>
		</tr>
		<tr>
			<td>DNase I</td>
			<td>5PRIMER</td>
			<td>2500120</td>
			<td></td>
		</tr>
		<tr>
			<td>isofluorane&#160;</td>
			<td>Halocarbon</td>
			<td>NPC12164-002-25</td>
			<td></td>
		</tr>
		<tr>
			<td>formaldehyde</td>
			<td>MACRON</td>
			<td>H121-08</td>
			<td></td>
		</tr>
		<tr>
			<td>EtOH (Ethanol)</td>
			<td>J.T. Baker</td>
			<td>800605</td>
			<td></td>
		</tr>
		<tr>
			<td>glacial acetic acid</td>
			<td>Panreac</td>
			<td>131008.1611</td>
			<td></td>
		</tr>
		<tr>
			<td>aluminum potassium sulfate</td>
			<td>Sigma-Aldrich</td>
			<td>12625</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene&#160;</td>
			<td>Leica</td>
			<td>3803665</td>
			<td></td>
		</tr>
		<tr>
			<td>0.22 &#956;m membranes</td>
			<td>Merck Millipore</td>
			<td>Millex-GP</td>
			<td></td>
		</tr>
		<tr>
			<td>AUTOCLIP Wound Clips, 9 mm</td>
			<td>BD Biosciences</td>
			<td>427631</td>
			<td></td>
		</tr>
		<tr>
			<td>AUTOCLIP Wound Clip Applier</td>
			<td>BD Biosciences</td>
			<td>427630</td>
			<td></td>
		</tr>
		<tr>
			<td>CellMask&#8482; Deep Red</td>
			<td>Thermo Fisher Scientific&#160;</td>
			<td>C10046</td>
			<td>plasma membrane stain</td>
		</tr>
	</tbody>
</table>

transfer of mammary gland-forming ability between mammary basal epithelial cells and mammary luminal cells via extracellular vesicles/exosomes

Le cellule possono comunicare attraverso esosomi, vescicole extracellulari di ~ 100 nm (EVs) che contengono proteine, lipidi e acidi nucleici. Le vescicole extracellulari ottenute dalle cellule epiteliali mammarie non mescolate (NAMEC) possono essere isolate dal mezzo NAMEC tramite ultracentrifugazione differenziale. Sulla base della loro densità, gli EV possono essere purificati mediante ultracentrifugazione a 110.000 x g. La preparazione EV da ultracentrifugazione può essere ulteriormente separata usando un gradiente continuo di densità per impedire la contaminazione con proteine ​​solubili. Gli EVs purificati possono quindi essere ulteriormente valutati usando un&#39;analisi di tracking nanoparticolare, che misura la dimensione e il numero di vescicole nel preparato. Le vescicole extracellulari con una dimensione variabile da 50 a 150 nm sono esosomi. Gli EVs / esosomi derivati ​​dal NAMEC possono essere ingeriti da cellule epiteliali mammarie, che possono essere misurate mediante citometria a flusso e microscopia confocale. Alcune proprietà delle cellule staminali del mammario ( ad esempio, la capacità di formare la ghiandola mammaria) possonoEssere trasferito da NAMEC-simili a gambo alle cellule epiteliali mammarie attraverso i EV / esosomi derivati ​​da NAMEC. Le cellule epiteliali mammarie luminose luminescenti epCAM hi / CD49f non possono formare ghiandole mammarie dopo essere state trapiantate nei rilievi di grasso del mouse, mentre le cellule epiteliali mammarie EpCAM lo / CD49f hi basali formano ghiandole mammarie dopo il trapianto. L&#39;assorbimento di EV / esosomi derivati ​​da NAMEC da EpCAM hi / CD49f lo luminal cellule epiteliali mammarie permette loro di generare ghiandole mammarie dopo essere stati trapiantati in pastiglie grasse. Gli EVs / esosomi derivati ​​da cellule epiteliali mammarie simili al gambo trasferiscono la capacità di formare la ghiandola mammaria alle cellule epiteliali mammarie luminose EpCAM hi / CD49f.

Dal momento che è stato dimostrato che bloccando la segnalazione di PGE 2 / EP 4 provoca il rilascio di EV / esosomi da cellule staminali simili al basale mammario 4 , questo lavoro presenta un metodo di isolamento degli EV / esosomi indotti dalla coltura cellulare basale epiteliale mammaria (NAMEC). Dal momento che NAMECs sono coltivati ​​in un mezzo senza siero, non esistono EVs / esosomi preesistenti derivati ​​dal siero <sup clas...

Watch this Scientific Journal Video about Transfer of Mammary Gland-forming Ability Between Mammary Basal Epithelial Cells and Mammary Luminal Cells via Extracellular Vesicles/Exosomes at JoVE.com

Transfer of Mammary Gland-forming Ability Between Mammary Basal Epithelial Cells and Mammary Luminal Cells via Extracellular Vesicles/Exosomes

Questo protocollo descrive metodi per purificare, quantificare e caratterizzare vescicole extracellulari (EVs) / esosomi da cellule epiteliali mammarie non aderenti / mesenchimali e per usarle per trasferire la capacità di formazione delle ghiandole mammarie alle cellule epiteliali mammarie luminali. EVs / esosomi derivati ​​da cellule epiteliali mammarie simili a stelo possono trasferire questa proprietà cellulare alle cellule che ingeriscono gli EV / esosomi.

Trasferimento di abilità di formazione del ghiandale mammario tra le cellule epiteliali basali del mammifero e le cellule luminescenti mammarie attraverso le vescicole / esosomi extracellulari

Cells can communicate via exosomes, ~100-nm extracellular vesicles (EVs) that contain proteins, lipids, and nucleic acids. Non-adherent/mesenchymal ...

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Cultivate Primary Nasal Epithelial Cells from Children and Reprogram into Induced Pluripotent Stem Cells

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