Name: Toepassing van een nieuwe hyaluronzuurhydrogel voor driedimensionale follikelcultuur en methodologie voor cryopreservatie van de ovariële follikel bij muizen (Video) | JoVE Journal
Uploaded: 2025-05-09T13:40:00.000+00:00
Duration: 10 min 31 s
Description: Leer een nieuwe hyaluronzuurhydrogelmethode voor driedimensionale follikelcultuur en cryopreservatietechnieken bij muizen. Dit model behoudt de 3D-architectuur voor oöcytematiging en heeft potentieel voor vruchtbaarheidsbehandelingen.

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<li>Wu, J., Nayudu, P. L., Kiesel, P. S., Michelmann, H. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Luteinizing+hormone+has+a+stage-limited+effect+on+preantral+follicle+development+in+vitro%5BTitle%5D)+AND+%22Biol+Reprod%22%5BJournal%5D)">Luteinizing hormone has a stage-limited effect on preantral follicle development in vitro.</a> Biol Reprod. 63 (1), 320-327 (2000).</li>
</ol>

<table><tbody><tr><td>Anti-alpha tubulin-FITC gelabeld</td><td>Sigma-Aldrich</td><td>F2168</td><td></td></tr><tr><td>Anti-beta tubulin-FITC gelabeld</td><td>Sigma-Aldrich</td><td>F2043</td><td></td></tr><tr><td>BZ-X700</td><td>Keyence</td><td></td><td></td></tr><tr><td>Center well dish</td><td>Fisher Scientific</td><td>08-772-12</td><td></td></tr><tr><td>Collageenase Type I</td><td>Worthington Biochemical Corporation</td><td>LS004196</td><td></td></tr><tr><td>Crycap vial-geventileerd</td><td>Hampton Research</td><td>HR4-904</td><td></td></tr><tr><td>Cryoloop</td><td>Hampton Research</td><td>HR4-974</td><td></td></tr><tr><td>Kristalkap</td><td>Hampton Research</td><td>HR4-733</td><td></td></tr><tr><td>Kweekschaal 60mm</td><td>Fisher Scientific&nbsp;</td><td>08-772B</td><td></td></tr><tr><td>Dimethyl sulfoxide (DMSO)</td><td>Sigma-Aldrich</td><td>D2650</td><td></td></tr><tr><td>Epidermale groeifactor (EGF)</td><td>R &amp;D Systems</td><td>236-EG</td><td></td></tr><tr><td>Ethyleenglycol (EG)</td><td>Sigma-Aldrich</td><td>293237</td><td></td></tr><tr><td>Foetaal runderserum-hitte geïnactiveerd</td><td>ThermoFisher Scientific</td><td>10082-147</td><td></td></tr><tr><td>Follikelstimulerend hormoon (FSH)</td><td>Sigma-Aldrich</td><td>F4021</td><td></td></tr><tr><td>Global-Hepes&nbsp; medium</td><td>CooperSurgical</td><td>LGGH-100</td><td></td></tr><tr><td>Hoechst 33342</td><td>Sigma-Aldrich</td><td>B2261</td><td></td></tr><tr><td>Human choriongonadotrofine (hCG)</td><td>Sigma-Aldrich</td><td>CG10</td><td></td></tr><tr><td>Humaan serumalbumine</td><td>CooperSurgical</td><td>GHSA-125</td><td></td></tr><tr><td>Hyaluronidase</td><td>CooperSurgical</td><td>ART-4007-A</td><td></td></tr><tr><td>Waterstofperoxide</td><td>CVS Pharmacy Inc.</td><td>372441</td><td></td></tr><tr><td>Insuline-transferrine-selenium (ITS)</td><td>ThermoFisher Scientific</td><td>41400-045</td><td></td></tr><tr><td>Leibovitz medium (L-15)</td><td>ThermoFisher Scientific</td><td>11415-064</td><td></td></tr><tr><td>Luteïniserend hormoon</td><td>Sigma-Aldrich</td><td>L9773</td><td></td></tr><tr><td>Magnetische stok</td><td>Hampton Research</td><td>HR4-729</td><td></td></tr><tr><td>Micropipetten (1000 &micro;m)</td><td>Minitube</td><td>19025/0050</td><td></td></tr><tr><td>Micropipetten (175, 200, en 275&micro;m)</td><td>CooperSurgical</td><td>MXL3-175, MXL3-200, MXL3-275</td><td></td></tr><tr><td>Millex GV filter 0.22 &micro;m</td><td>Millipore</td><td>SLGU033RS</td><td></td></tr><tr><td>Mineralolie</td><td>CooperSurgical</td><td>LGOL-500</td><td></td></tr><tr><td>Minimum Essential Medium alpha (MEM)</td><td>ThermoFisher Scientific</td><td>32561-037</td><td></td></tr><tr><td>Oöcyten beeldvormingssysteem-Spindleview</td><td>Hamilton Thorne</td><td></td><td></td></tr><tr><td>Fosfaat gebufferde zoutoplossing (PBS)</td><td>ThermoFisher Scientific</td><td>10010-023</td><td></td></tr><tr><td>Propidium jodide</td><td>Sigma-Aldrich</td><td>P4170</td><td></td></tr><tr><td>Rapid i</td><td>VitroLife</td><td>14406</td><td></td></tr><tr><td>SmartBox</td><td>VitroLife</td><td>14423</td><td></td></tr><tr><td>Synthetisch eiwitsubstituut (SPS)</td><td>CooperSurgical</td><td>ART-3011</td><td></td></tr><tr><td>Tyramine -gekoppeld Hyaluronan Biohydrogel Kit&nbsp;</td><td>LifeCore</td><td>ENG-00151</td><td></td></tr><tr><td>Ultrasone sealer</td><td>VitroLife</td><td>14415</td><td></td></tr><tr><td>Universele GPS kweekschaal 8x 100 &micro;l putjes</td><td>CooperSurgical</td><td>UGPS-010</td><td></td></tr></tbody></table>

toepassing van een nieuwe hyaluronzuurhydrogel voor driedimensionale follikelcultuur en methodologie voor cryopreservatie van de ovariële follikel bij muizen

Application of a Novel Hyaluronan Hydrogel for Three-Dimensional Follicle Culture and Methodology for Mouse Ovarian Follicle Cryopreservation

Toepassing van een nieuwe hyaluronzuurhydrogel voor driedimensionale follikelcultuur en methodologie voor cryopreservatie van de ovariële follikel bij muizen

The 3-D architecture of the ovarian follicle and the complex interactions between somatic cell components and the oocyte that are necessary for ...

application-novel-hyaluronan-hydrogel-for-three-dimensional-follicle

Research

JoVE Journal

Biology

Biologie

Een nieuwe methode voor ovariële follikel inkapseling in een 3D-fibrine-alginaat interpenetrerend netwerk wordt beschreven. Dit systeem combineert een structurele ondersteuning met proteolytische degradatie van de ontwikkeling van onvolgroeide follikels steun aan rijpe eicellen te produceren. Deze methode kan worden toegepast op de cultuur cel aggregaten tot cel-cel contacten, zonder beperking van de uitbreiding te behouden.

Een methode voor ovariële follikel Encapsulation en cultuur in een proteolytisch Afbreekbare 3 dimensionale systeem

Cultuur van normale cellen in hun drie-dimensionale context is een alternatieve methode voor vroege gebeurtenissen die nodig is voor cellulaire transformatie en tumorvorming te bestuderen. Deze methode wordt gebruikt om de normale eierstokken en oviductal cellen groeien naar vroege gebeurtenissen bij eierstokkanker vorming te bestuderen.

Alginaat Hydrogelen voor Three-Dimensional orgelcultuur van de eierstokken en eileiders

Dit protocol beschrijft de primaire cultuur / co-cultuur van de muis eierstokweefsel, met behulp van de eierstokken van pasgeboren muizen en individuele ovariële follikels van prepuberale muizen. De kweektechnieken ondersteunen In een sterk fysiologische wijze, waardoor onderzoek naar het effect van extrinsieke middelen op de eierstok en interacties tussen ovariële follikels.

Cultuur en co-cultuur van de Muis eierstokken en ovariële follikels

Een zeer veelbelovende techniek voor het genereren van weefsel construeert zonder met behulp van matrix aan cultuur cellen in een gesimuleerde microzwaartekracht voorwaarde is. Met behulp van een roterende cultuur systeem, onderzochten we ovariële follikel groei en oöcyt rijping in termen van overleving van de follikel, morfologie, groei en oöcyt functie onder de voorwaarde van de gesimuleerde microzwaartekracht.

In Vitro Groei van de Preantral follikels muis onder gesimuleerde microzwaartekracht

Ovariële weefselcultures kan worden gebruikt als modellen van follikel ontwikkeling, ovulatie en follikel atresie en regulerende mechanismen van dynamische ovariële processen aan te geven.

Ovariële weefselkweek te visualiseren verschijnselen in muis eierstok

Hier worden twee protocollen gepresenteerd voor de inkapseling van varkensocyten in 3D-kweekomstandigheden. In de eerste worden cumulus-eicellencomplexen (COC's) ingekapseld in fibrin-alginaatkralen. In de tweede zijn ze ingesloten met gefluoreerde ethyleenpropyleenpoederdeeltjes (microbioreactoren). Beide systemen zorgen voor optimale omstandigheden om hun 3D-organisatie te onderhouden.

Proteolytisch afgebroken alginaat hydrogels en hydrofobe microbioreactoren voor porcine Oocyte Inkapseling

Hier beschrijven we een protocol dat beschrijft hoe menselijke ovariële follikels kunnen worden geïsoleerd uit bevroren ontdooid corticale weefsel om genexpressieanalyses uit te voeren.

Genexpressieanalyses in menselijke follikels

We beschrijven methoden om ovariële follikels te kweken in een nieuwe steigervrije agaroseschimmel die een aanvulling vormt op in vitro gametogenese.

Productie en gebruik van aanpasbare agarose-mallen voor steigervrije ovariële follikelcultuur van muizen

Beschrijven we een chip-gebaseerde platform voor de drie-dimensionale kweken van cellen in micro-bioreactoren. Een chip kan maximaal huis tot 10 miljoen. cellen die kunnen worden geteeld onder nauwkeurig omschreven voorwaarden met betrekking tot stroming, zuurstofspanning enz. in een steriele, gesloten circulatie lus.

Chip-op basis van drie-dimensionale Cultuur van de Cel in doorbloed Micro-bioreactoren

We presenteren twee processen voor de microfabricage van poreuze polymeer chips voor drie-dimensionale celkweek. De eerste is hot embossing gecombineerd met een damp van oplosmiddelen lasproces. De tweede maakt gebruik van een recent ontwikkelde microthermoforming proces in combinatie met ion-Track technologie leidt tot een aanzienlijke vereenvoudiging van de productie.

Microfabricage van Chip-en kleinbedrijf Stellingen voor drie-dimensionale celkweek

Leer een nieuwe hyaluronzuurhydrogelmethode voor driedimensionale follikelcultuur en cryopreservatietechnieken bij muizen. Dit model behoudt de 3D-architectuur voor oöcytematiging en heeft potentieel voor vruchtbaarheidsbehandelingen.

Toepassing van een nieuwe hyaluronzuurhydrogel voor driedimensionale follikelcultuur en methodologie voor cryopreservatie van de ovariële follikel bij muizen (Video) | JoVE Journal

Toepassing van een nieuwe hyaluronzuurhydrogel voor driedimensionale follikelcultuur en methodologie voor cryopreservatie van de ovariële follikel bij muizen

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