Name: Rilevamento della senescenza utilizzando luce riflessa nella frazione vascolare stromale adiposa (Video) | JoVE Journal
Uploaded: 2026-06-05T13:15:00.000+00:00
Duration: 10 min 8 s
Description: Impara a rilevare la senescenza nelle cellule SVF umane mediante microscopia confocale a luce riflessa. Questo metodo quantitativo consente analisi single-cell oggettiva e immunocitochimica multiplex per marcatori di senescenza e lignaggio.

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</ol>

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rilevamento della senescenza utilizzando luce riflessa nella frazione vascolare stromale adiposa

Senescence Detection Using Reflected Light in Adipose Stromal Vascular Fraction

Rilevamento della senescenza utilizzando luce riflessa nella frazione vascolare stromale adiposa

Cellular senescence is a stress-induced state characterized by permanent cell-cycle arrest and the development of a distinctive secretory profile that ...

senescence-detection-using-reflected-light-adipose-stromal-vascular

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Biologia

Metodi tradizionali di valutazione adipogenico differenziazione sono economici e facili da usare, ma non sono specifici di cambiamenti nell'espressione genica. Abbiamo sviluppato un test per quantificare la differenziazione delle cellule mesenchimali in adipociti maturi con un pennarello specifico lignaggio. Questo test ha diverse applicazioni attraverso la ricerca di base e della medicina clinica.

Visualizzazione e quantificazione del potenziale di differenziazione delle cellule mesenchimali adipogenico con un Marker specifico lignaggio

A causa del contenuto elevato del lipido, tessuto adiposo è stato impegnativo per visualizzare usando i metodi istologici tradizionali. Adipo-Clear è un tessuto tecnica che permette di etichettatura robusto e imaging fluorescente volumetrica ad alta risoluzione del tessuto adiposo di compensazione. Qui, descriviamo i metodi per la preparazione del campione, pretrattamento, colorazione, compensazione e montaggio per l'imaging.

Adipo-Clear: Un tessuto metodo di compensazione per l'Imaging tridimensionale del tessuto adiposo

L'obiettivo del presente studio è di dimostrare la tecnica immunostaining e visualizzazione di intero-monta come un metodo ideale per l'imaging 3D della componente cellulare e architettura del tessuto adiposo.

Visualizzazione della struttura del tessuto adiposo bianco 3D usando la macchiatura del intero-supporto

Formazione del vaso sanguigno di nuovo e lo stimolo simpatico svolgono un ruolo fondamentale nel rimodellamento del tessuto adiposo. Tuttavia, permangono problemi tecnici nella visualizzazione e quantitativamente misurando il tessuto adiposo. Qui presentiamo un protocollo per etichettare correttamente e confrontare quantitativamente le densità dei vasi sanguigni e fibre nervose in diversi tessuti adiposi.

Co-macchiatura dei vasi sanguigni e fibre nervose nel tessuto adiposo

Questo protocollo fornisce un metodo efficiente di digestione della collagenasi per l'isolamento di adipociti vitali e cellule di frazione vascolare stromale-SVF dal grasso viscerale umano in un unico processo, compresa la metodologia per ottenere RNA di alta qualità dagli adipociti e la fenotipizzazione dei macrofagi SVF attraverso la colorazione di più marcatori legati alla membrana per l'analisi mediante citometria a flusso.

Isolamento di adipociti vitali e frazione vascolare stromale dal tessuto adiposo viscerale umano adatto per l'analisi dell'RNA e la fenotipizzazione dei macrofagi

I preadipociti sono isolati dalla frazione vascolare stromale del tessuto adiposo bruno interscapolare di topi neonati e differenziati in cellule che accumulano goccioline lipidiche, esprimono marcatori molecolari e mostrano la morfologia mitocondriale degli adipociti bruni maturi. Queste cellule vengono ulteriormente analizzate mediante immunofluorescenza e microscopia elettronica a trasmissione.

Differenziazione e imaging di adipociti marroni dalla frazione vascolare stromale del tessuto adiposo interscapolare da topi neonati

Questo protocollo descrive una metodologia per isolare e identificare le cellule staminali mesenchimali (MSC) derivate dal tessuto adiposo da ratti Sprague Dawley.

Isolamento e identificazione di cellule staminali mesenchimali derivate dal tessuto adiposo di ratti Sprague Dawley

Questo protocollo descrive l'isolamento semi-automatico della frazione vascolare stromale (SVF) dal tessuto adiposo murino per ottenere preadipociti e ottenere la differenziazione degli adipociti in vitro. L'uso di un dissociatore tissutale per la digestione della collagenasi riduce la variazione sperimentale e aumenta la riproducibilità.

Isolamento semi-automatico della frazione vascolare stromale dal tessuto adiposo bianco murino mediante un dissociatore tissutale

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Studio del cross-talk tra cellule endoteliali adipose e adipociti nel tessuto adiposo sottocutaneo umano

Qui presentiamo un metodo standard per isolare in modo efficiente cellule staminali e adipociti derivati dalle adipoci, caratterizzato da una riduzione del &gt;90% dei tempi di lavorazione, un'elevata vitalità cellulare e una compatibilità ampia.

Isolamento efficiente di cellule staminali e adipociti derivati dall'adiposo dal tessuto adiposo suino

Impara a rilevare la senescenza nelle cellule SVF umane mediante microscopia confocale a luce riflessa. Questo metodo quantitativo consente analisi single-cell oggettiva e immunocitochimica multiplex per marcatori di senescenza e lignaggio.

Rilevamento della senescenza utilizzando luce riflessa nella frazione vascolare stromale adiposa

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