Name: an ex vivo model to study hormone action in the human breast
Uploaded: 2015-01-08T15:00:00
Description: Watch this Scientific Journal Video about An Ex vivo Model to Study Hormone Action in the Human Breast at JoVE.com

Forfatterne takker M. Fiche af University Hospital of Lausanne for at give mammoplasty fotografi, M. Wirth og A. Ayyanan af Swiss Institute for Experimental Cancer Research, Det Nationale Center for Kompetence i forskning Molecular Oncology, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne til faglig bistand og R. Clarke fra University of Manchester til kritiske bemærkninger. Den forskning, der fører til disse resultater har fået støtte fra SNF3100A0-112090, Oncosuisse 531.817, og initiativet om innovative lægemidler fællesforetagende i henhold tilskudsaftale nej. 115.188, ressourcer der består af finansielle bidrag fra Den Europæiske Unions syvende rammeprogram (FP7 / 2007-2013) og European Federation of Pharmaceutical Industries og foreninger virksomheder «i naturalier bidrag. Webadressen på initiativet om innovative lægemidler er <a href="http://www.imi.europa.eu/" target="_blank">http://www.imi.europa.eu/</a> . &lt;/ P&gt;

Generelle betragtninger: i god tid, der er nedsat en etisk protokol, forberede spørgeskemaer til patienter, og se til at uddanne de kliniske personale. Før du bruger materiale fra reduktion mammoplasty kirurgi, fremskaffe tilladelser, og sikre patientens samtykke. Væv kan være smittefarlige og skal behandles i overensstemmelse hermed. Denne protokol blev godkendt af den etiske komité i ISREC - Swiss Institute for Experimental Cancer Research. 1. Tissue Recovery, forarbejdning og Bio-banki...

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J., Echeverria, P., Brisken, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ER+and+PR+signaling+nodes+during+mammary+gland+development.">ER and PR signaling nodes during mammary gland development.</a> Breast Cancer Res. 14 (4), 210 (2012).</li><li>Roskelley, C., Desprez, P., Bissell, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+matrix-dependent+tissue-specific+gene+expression+in+mammary+epithelial+cells+requires+both+physical+and+biochemical+signal+transduction.">Extracellular matrix-dependent tissue-specific gene expression in mammary epithelial cells requires both physical and biochemical signal transduction.</a> Proc Natl Acad Sci U S A. 91 (26), 12378-12382 (1994).</li><li>Kass, L., Erler, J. T., Dembo, M., Weaver, V. 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Den ex vivo kultur beskrevet her tilvejebringer brystvæv mikrostrukturer indeholdende intakte kanaler og lobules, sammen med andre celletyper, der normalt findes i den humane kvindelige bryst. I behandlingen af ​​brystvæv menneske normalt fra reduktion mammoplasties, fjernelse af fedtvæv, mekanisk og enzymatisk fordøjelse af stromale matrix, og lyse af røde blodlegemer beriger for mælk ventilationskanal fragmenter og terminale duktale luftrør enheder. Gentle enzymatisk nedbrydning på det rigtige kon...

A correction was made to <a href="/video/52436">An Ex Vivo Model to Study Hormone Action in the Human Breast</a>. There was an error with the author, Marie Shamsheddin&#39;s, name. The author&#39;s name has been corrected to:
Marie Shamseddin
instead of:
Marie Shamsheddin

A correction to the author list has been made for the article <a href="/video/52436">An Ex Vivo Model to Study Hormone Action in the Human Breast</a>.

Erratum: An Ex Vivo Model to Study Hormone Action in the Human Breast

Oplysninger om den mutational landskab i brystkræft er stigende på hurtigt tempo. Mindre opmærksomhed er blevet betalt til systemiske faktorer, der påvirker brystkræft udvikling. Udsættelse for reproduktive hormoner har stor indflydelse på sygdomsudvikling 1-3. Alligevel er de mekanismer, som reproduktive hormoner kolliderer med menneskelige bryst dårligt forstået. Arbejde med gensplejsede musemodeller har vist, at de involverer celle iboende og parakrin signalering gennem flere nedstrømseffektorer 4. Den begrænsede viden om hormon handling i den menneskelige bryst skyldes i høj grad mangel på hensigtsmæssige modeller. Meste arbejde om mekanismerne af østrogen-receptor (ER) og progesteron-receptor (PR) signalering er blevet udført med hormonreceptor positive brystkræft cellelinier, såsom MCF-7 og T47D. Disse blev afledt af pleuraekssudat fra patienter med fremskreden brystcancer, som havde already fik flere behandlinger 5. Den biologiske relevans af resultaterne i sådanne enkle in vitro modeller af den menneskelige bryst er tvivlsomme og målgener identificeret i disse in vitro modeller er forskellige fra målgener, der er identificeret i dyremodeller 6. Når primære humane bryst epiteliale celler dyrkes in vitro de tendens til at miste hormon receptor-ekspression og dermed hormon respons 7,8. Dette problem kan circumventd af avancerede 3D-metoder, der anvender matrigel. På denne måde lykkedes C. Clarke og kolleger oprettelse bryst epiteliale celler, opretholdt hormon receptor-ekspression og viste en proliferativ reaktion på progesteron stimulation 9. Men to vigtige in vivo progesteron receptor målgener, Wnt-4 og RANKL, blev ikke induceret ved progesteron stimulation i dette system 9. Denne fremgangsmåde blev for nylig taget på videre med in vitro </em&gt; hormon forbehandling og RANKL induktion blev opnået 10. En advarsel er, at matrigel har aktiviteter, der er batch-afhængig, er dyre, og kræver et eksperimentelt design, der er velegnet til små celle kun tal. Baseret på den konstatering, at in vivo-ER og PR-signalering er i høj grad medieret af parakrine interaktioner 11, fremførte vi, at der skal opretholdes intercellulære interaktioner. En anden vigtig faktor, der er tabt som væv dissocieret til enkelte celler til in vitro kultur vekselvirkninger epitelceller med den ekstracellulære matrix; endnu disse er kritiske for epitel differentiering og deres forstyrrelser er vigtig i tumorgenese 12. Med dette i tankerne, vi etableret en metode til at isolere bryst væv mikrostrukturer fra frisk kirurgisk udsmid materiale 13. Bryst parenkym Den består af en to-lags epitel med indvendig luminale og ydre myoepitheliAL-celler, dissekeres væk fra fedtvæv og udsættes for mekanisk og enzymatisk dissociation. Efter vask og centrifugering, er fragmenter af mælkekanaler opnået som bevarer nære interaktioner med mange stromaceller. Disse væv mikrostrukturer forblive hormon lydhør. Modellen blev valideret i kliniske prøver 13. Som sådan kan den nuværende procedure med til at studere hormonvirkning i brystet i en biologisk og klinisk relevant sammenhæng.

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an ex vivo model to study hormone action in the human breast

Studiet af hormonvirkning i humane bryst- er blevet hæmmet af manglende passende modelsystemer. Ved in vitro kultur, primære mammaepitelceller tendens til at miste hormon receptor ekspression. Udbredte hormon receptor positiv brystkræft cellelinier er af begrænset relevans for in vivo situation. Her beskriver vi en ex vivo model til at studere hormon indsats i human bryst. Friske humane bryst vævsprøver fra kirurgisk udsmid materiale såsom mammoplasties eller mammectomies reduktion er mekanisk og enzymatisk fordøjet at opnå vævsfragmenter indeholder kanaler og lobules og flere stromale celletyper. Disse væv mikrostrukturer holdes i basalt medium uden vækstfaktorer bevare deres intercellulære kontakter, vævet arkitektur, og forblive hormon lydhør i flere dage. De er let behandles til RNA og protein ekstraktion, histologisk analyse eller opbevares i frysemedium. Fluorescenscellesortering (FACS) kan anvendes til at berige for specifikke cellepopulationer. Denne protokol giver en ligefrem, standard metode til translationelle studier med meget komplekse, varierede humane prøver.

For at undersøge den rolle, østrogener og progesteron og for bedre at forstå deres molekylære funktioner i den menneskelige bryst, vi indsamler friske humane bryst vævsprøver fra patienter, der gennemgår reduktion mammoplasties (figur 1) efter at have opnået deres informerede samtykke. Vi har også opnå patienters og reproduktiv historie samt en blodprøve for at bestemme serum progesteron niveauer på tidspunktet for kirurgi. Væv fra friske reduktion mammoplasties er mekanisk og enzymatisk di...

Watch this Scientific Journal Video about An Ex vivo Model to Study Hormone Action in the Human Breast at JoVE.com

An Ex vivo Model to Study Hormone Action in the Human Breast

Vi har udviklet en ny ex vivo-model til at studere hormon indsats i human bryst. Den er baseret på væv mikrostrukturer isoleret fra kirurgisk bryst vævsprøver, som bevarer væv arkitektur, intercellulære interaktioner, og parakrin signalering.

En<em&gt; Ex vivo</em&gt; Model til Uddannelse hormonvirkning i Human Breast

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial ...

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An Ex vivo Model to Study Hormone Action in the Human Breast

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Organ fibrosis or &#8220;scarring&#8221; is known to account for a high death toll due to the extensive amount of disorders and organs affected (from ...

Methods for Culturing Human Femur Tissue Explants to Study Breast Cancer Cell Colonization of the Metastatic Niche

Metoder til dyrkning menneskelige femur vævseksplantater at studere Breast Cancer Cell kolonisering af metastatisk Niche

Bone is the most common site of breast cancer metastasis. Although it is widely accepted that the microenvironment influences cancer cell behavior, little ...

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

In vivo og ex vivo Approaches at studere kræft i æggestokkene Metastatisk Kolonisering af Milky Spot Strukturer i peritoneal Adipose

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of ...

An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis

En ortotopisk Mouse Model af Spontan Breast Cancer Metastase

Metastasis is the primary cause of mortality of breast cancer patients. The mechanism underlying cancer cell metastasis, including breast cancer ...

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

Udtømning af Mouse Celler fra Humane tumorxenoplantater Markant Forbedrer Downstream Analyse af Target Cells

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

Breast Milk Enhances Growth of Enteroids: An Ex Vivo Model of Cell Proliferation

Breast Milk Enhances Growth of Enteroids: An Ex Vivo Model of Cell Proliferation

Modermælk øger væksten af Enteroids: en Ex Vivo Model af celledelingen

Human small intestinal enteroids are derived from the crypts and when grown in a stem cell niche contain all of the epithelial cell types. The ability to ...