Name: Innesti adiposi bianchi e marroni: un approccio per correggere i deficit riproduttivi, metabolici e renali nei topi obesi Black and Tan Brachyury (BTBR) (Video) | JoVE Journal
Uploaded: 2025-09-09T13:45:00.000+00:00
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Description: Scopri il trapianto combinato di tessuto adiposo bianco e bruno per correggere disfunzioni metaboliche e riproduttive nei topi obesi BTBR. Questo protocollo standardizzato migliora la fertilità, il metabolismo e la funzione renale.

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<ol><li>Zhao, M., Cao, Y., Ma, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+insights+in+the+treatment+of+DKD%3A+recent+advances+and+future+prospects%5BTitle%5D)+AND+%22BMC+Nephrol%22%5BJournal%5D)">New insights in the treatment of DKD: recent advances and future prospects.</a> BMC Nephrol. 26 (1), 72(2025).</li>
<li>van Raalte, D. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Combination+therapy+for+kidney+disease+in+people+with+diabetes+mellitus%5BTitle%5D)+AND+%22Nat+Rev+Nephrol%22%5BJournal%5D)">Combination therapy for kidney disease in people with diabetes mellitus.</a> Nat Rev Nephrol. 20 (7), 433-446 (2024).</li>
<li>Xu, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Trends+and+future+predictions+of+chronic+kidney+disease+due+to+diabetes+mellitus+type+2+attributable+to+dietary+risks%3A+insights+based+on+GBD+2021+data%5BTitle%5D)+AND+%22Front+Nutr%22%5BJournal%5D)">Trends and future predictions of chronic kidney disease due to diabetes mellitus type 2 attributable to dietary risks: insights based on GBD 2021 data.</a> Front Nutr. 11, 1494383(2024).</li>
<li>GBD 2021 Adult BMI Collaborators. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adult+BMI+Collaborators.+Global%2C+regional%2C+and+national+prevalence+of+adult+overweight+and+obesity%2C+1990-2021%2C+with+forecasts+to+2050%3A+a+forecasting+study+for+the+Global+Burden+of+Disease+Study+2021%5BTitle%5D)+AND+%22Lancet%22%5BJournal%5D)">Adult BMI Collaborators. Global, regional, and national prevalence of adult overweight and obesity, 1990-2021, with forecasts to 2050: a forecasting study for the Global Burden of Disease Study 2021.</a> Lancet. 405 (10481), 813-838 (2025).</li>
<li>Liu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Brown+adipose+tissue+transplantation+reverses+obesity+in+Ob%2FOb+mice%5BTitle%5D)+AND+%22Endocrinology%22%5BJournal%5D)">Brown adipose tissue transplantation reverses obesity in Ob/Ob mice.</a> Endocrinology. 156 (7), 2461-2469 (2015).</li>
<li>Ghesmati, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+update+on+the+secretory+functions+of+brown%2C+white%2C+and+beige+adipose+tissue%3A+Towards+therapeutic+applications%5BTitle%5D)+AND+%22Rev+Endocr+Metab+Disord%22%5BJournal%5D)">An update on the secretory functions of brown, white, and beige adipose tissue: Towards therapeutic applications.</a> Rev Endocr Metab Disord. 25 (2), 279-308 (2024).</li>
<li>Dewal, R. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Transplantation+of+committed+pre-adipocytes+from+brown+adipose+tissue+improves+whole-body+glucose+homeostasis%5BTitle%5D)+AND+%22iScience%22%5BJournal%5D)">Transplantation of committed pre-adipocytes from brown adipose tissue improves whole-body glucose homeostasis.</a> iScience. 27 (2), 108927(2024).</li>
<li>Klebanov, S., Astle, C. M., DeSimone, O., Ablamunits, V., Harrison, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adipose+tissue+transplantation+protects+ob%2Fob+mice+from+obesity%2C+normalizes+insulin+sensitivity+and+restores+fertility%5BTitle%5D)+AND+%22J+Endocrinol%22%5BJournal%5D)">Adipose tissue transplantation protects ob/ob mice from obesity, normalizes insulin sensitivity and restores fertility.</a> J Endocrinol. 186 (1), 203-211 (2005).</li>
<li>Childs, G. V., Odle, A. K., MacNicol, M. C., MacNicol, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+importance+of+leptin+to+reproduction%5BTitle%5D)+AND+%22Endocrinology%22%5BJournal%5D)">The importance of leptin to reproduction.</a> Endocrinology. 162 (2), 1-18 (2021).</li>
<li>Hudkins, K. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((BTBR+Ob%2FOb+mutant+mice+model+progressive+diabetic+nephropathy%5BTitle%5D)+AND+%22J+Am+Soc+Nephrol%22%5BJournal%5D)">BTBR Ob/Ob mutant mice model progressive diabetic nephropathy.</a> J Am Soc Nephrol. 21 (9), 1533-1542 (2010).</li>
<li>Stanford, K. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Brown+adipose+tissue+regulates+glucose+homeostasis+and+insulin+sensitivity%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">Brown adipose tissue regulates glucose homeostasis and insulin sensitivity.</a> J Clin Invest. 123 (1), 215-223 (2013).</li>
<li>Pinckard, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+endocrine+role+for+the+BAT-released+Lipokine+12%2C13-diHOME+to+mediate+cardiac+function%5BTitle%5D)+AND+%22Circulation%22%5BJournal%5D)">A novel endocrine role for the BAT-released Lipokine 12,13-diHOME to mediate cardiac function.</a> Circulation. 143 (2), 145-159 (2021).</li>
<li>Valgas da Silva, P. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Brown+adipose+tissue+prevents+glucose+intolerance+and+cardiac+remodeling+in+high-fat-fed+mice+after+a+mild+myocardial+infarction%5BTitle%5D)+AND+%22Int+J+Obes+%28Lond%29%22%5BJournal%5D)">Brown adipose tissue prevents glucose intolerance and cardiac remodeling in high-fat-fed mice after a mild myocardial infarction.</a> Int J Obes (Lond). 46 (2), 350-358 (2022).</li>
<li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Brown+adipose+tissue+transplantation+ameliorates+diabetic+nephropathy+through+the+miR-30b+pathway+by+targeting+Runx1%5BTitle%5D)+AND+%22Metabolism%22%5BJournal%5D)">Brown adipose tissue transplantation ameliorates diabetic nephropathy through the miR-30b pathway by targeting Runx1.</a> Metabolism. 125, 154916(2021).</li>
<li>Clee, S. M., Nadler, S. T., Attie, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+and+genomic+studies+of+the+BTBR+ob%2Fob+mouse+model+of+type+2+diabetes%5BTitle%5D)+AND+%22Am+J+Ther%22%5BJournal%5D)">Genetic and genomic studies of the BTBR ob/ob mouse model of type 2 diabetes.</a> Am J Ther. 12 (6), 491-498 (2005).</li>
<li>Stoehr, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Identification+of+major+quantitative+trait+loci+controlling+body+weight+variation+in+ob%2Fob+mice%5BTitle%5D)+AND+%22Diabetes%22%5BJournal%5D)">Identification of major quantitative trait loci controlling body weight variation in ob/ob mice.</a> Diabetes. 53 (1), 245-249 (2004).</li>
<li>Liu, X., Zhang, Z., Song, Y., Xie, H., Dong, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+update+on+brown+adipose+tissue+and+obesity+intervention%3A+Function%2C+regulation+and+therapeutic+implications%5BTitle%5D)+AND+%22Front+Endocrinol+%28Lausanne%29%22%5BJournal%5D)">An update on brown adipose tissue and obesity intervention: Function, regulation and therapeutic implications.</a> Front Endocrinol (Lausanne). 13, 1065263(2022).</li>
<li>Mollah, M. L., Yang, H. S., Jeon, S., Kim, K., Cheon, Y. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Overaccumulation+of+fat+caused+rapid+reproductive+senescence+but+not+loss+of+ovarian+reserve+in+ob%2Fob+mice%5BTitle%5D)+AND+%22J+Endocr+Soc%22%5BJournal%5D)">Overaccumulation of fat caused rapid reproductive senescence but not loss of ovarian reserve in ob/ob mice.</a> J Endocr Soc. 5 (1), bvaa168(2021).</li>
<li>Serke, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin-deficient+%28ob%2Fob%29+mouse+ovaries+show+fatty+degeneration%2C+enhanced+apoptosis+and+decreased+expression+of+steroidogenic+acute+regulatory+enzyme%5BTitle%5D)+AND+%22Int+J+Obes+%28Lond%29%22%5BJournal%5D)">Leptin-deficient (ob/ob) mouse ovaries show fatty degeneration, enhanced apoptosis and decreased expression of steroidogenic acute regulatory enzyme.</a> Int J Obes (Lond). 36 (8), 1047-1053 (2012).</li>
<li>Diamanti-Kandarakis, E., Dunaif, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Insulin+resistance+and+the+polycystic+ovary+syndrome+revisited%3A+an+update+on+mechanisms+and+implications%5BTitle%5D)+AND+%22Endocr+Rev%22%5BJournal%5D)">Insulin resistance and the polycystic ovary syndrome revisited: an update on mechanisms and implications.</a> Endocr Rev. 33 (6), 981-1030 (2012).</li>
<li>Chang, A. S., Dale, A. N., Moley, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Maternal+diabetes+adversely+affects+preovulatory+oocyte+maturation%2C+development%2C+and+granulosa+cell+apoptosis%5BTitle%5D)+AND+%22Endocrinology%22%5BJournal%5D)">Maternal diabetes adversely affects preovulatory oocyte maturation, development, and granulosa cell apoptosis.</a> Endocrinology. 146 (5), 2445-2453 (2005).</li>
<li>Almog, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin+attenuates+follicular+apoptosis+and+accelerates+the+onset+of+puberty+in+immature+rats%5BTitle%5D)+AND+%22Mol+Cell+Endocrinol%22%5BJournal%5D)">Leptin attenuates follicular apoptosis and accelerates the onset of puberty in immature rats.</a> Mol Cell Endocrinol. 183 (1-2), 179-191 (2001).</li>
<li>Beyerstedt, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Combining+sodium-glucose+co-transporter-2+inhibitor+with+mesenchymal+stem+cells+and+brown+adipose+tissue+%28BAT%29+and+white+adipose+tissue+%28WAT%29+transplantation+to+mitigate+the+progression+of+diabetic+kidney+disease%3A+a+pre-clinical+approach%5BTitle%5D)+AND+%22Stem+Cell+Res+Ther%22%5BJournal%5D)">Combining sodium-glucose co-transporter-2 inhibitor with mesenchymal stem cells and brown adipose tissue (BAT) and white adipose tissue (WAT) transplantation to mitigate the progression of diabetic kidney disease: a pre-clinical approach.</a> Stem Cell Res Ther. 16 (254), 1-15 (2025).</li>
<li>Sávio-Silva, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Therapeutic+potential+of+mesenchymal+stem+cells+in+a+pre-clinical+model+of+diabetic+kidney+disease+and+obesity%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Therapeutic potential of mesenchymal stem cells in a pre-clinical model of diabetic kidney disease and obesity.</a> Int J Mol Sci. 22 (4), 1546(2021).</li>
<li>Takemura, S., Shimizu, T., Oka, M., Sekiya, S., Babazono, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Transplantation+of+adipose-derived+mesenchymal+stem+cell+sheets+directly+into+the+kidney+suppresses+the+progression+of+renal+injury+in+a+diabetic+nephropathy+rat+model%5BTitle%5D)+AND+%22J+Diabetes+Investig%22%5BJournal%5D)">Transplantation of adipose-derived mesenchymal stem cell sheets directly into the kidney suppresses the progression of renal injury in a diabetic nephropathy rat model.</a> J Diabetes Investig. 11 (3), 545-553 (2020).</li>
<li>Pichaiwong, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reversibility+of+structural+and+functional+damage+in+a+model+of+advanced+diabetic+nephropathy%5BTitle%5D)+AND+%22J+Am+Soc+Nephrol%22%5BJournal%5D)">Reversibility of structural and functional damage in a model of advanced diabetic nephropathy.</a> J Am Soc Nephrol. 24 (7), 1088-1102 (2013).</li>
<li>Almog, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin+attenuates+follicular+apoptosis+and+accelerates+the+onset+of+puberty+in+immature+rats%5BTitle%5D)+AND+%22Mol+Cell+Endocrinol%22%5BJournal%5D)">Leptin attenuates follicular apoptosis and accelerates the onset of puberty in immature rats.</a> Mol Cell Endocrinol. 183 (1-2), 179-191 (2001).</li>
<li>O'Brien, P. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((BTBR+ob%2Fob+mice+as+a+novel+diabetic+neuropathy+model%3A+Neurological+characterization+and+gene+expression+analysis%5BTitle%5D)+AND+%22Neurobiol+Dis%22%5BJournal%5D)">BTBR ob/ob mice as a novel diabetic neuropathy model: Neurological characterization and gene expression analysis.</a> Neurobiol Dis. 73, 348-355 (2015).</li>
<li>Sávio-Silva, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mesenchymal+stem+cell+therapy+for+diabetic+kidney+disease%3A+A+review+of+the+studies+using+syngeneic%2C+autologous%2C+allogenic%2C+and+xenogenic+cells%5BTitle%5D)+AND+%22Stem+Cells+Int%22%5BJournal%5D)">Mesenchymal stem cell therapy for diabetic kidney disease: A review of the studies using syngeneic, autologous, allogenic, and xenogenic cells.</a> Stem Cells Int. 2020, 8833725(2020).</li>
<li>Morsy, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin+gene+therapy+and+daily+protein+administration%3A+a+comparative+study+in+the+ob%2Fob+mouse%5BTitle%5D)+AND+%22Gene+Ther%22%5BJournal%5D)">Leptin gene therapy and daily protein administration: a comparative study in the ob/ob mouse.</a> Gene Ther. 5 (1), 8-18 (1998).</li>
<li>Wang, Y., Asakawa, A., Inui, A., Kosai, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin+gene+therapy+in+the+fight+against+diabetes%5BTitle%5D)+AND+%22Expert+Opin+Biol+Ther%22%5BJournal%5D)">Leptin gene therapy in the fight against diabetes.</a> Expert Opin Biol Ther. 10 (10), 1405-1414 (2010).</li>
<li>Blüher, S., Shah, S., Mantzoros, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin+deficiency%3A+clinical+implications+and+opportunities+for+therapeutic+interventions%5BTitle%5D)+AND+%22J+Investig+Med%22%5BJournal%5D)">Leptin deficiency: clinical implications and opportunities for therapeutic interventions.</a> J Investig Med. 57 (7), 784-788 (2009).</li>
<li>Paz-Filho, G., Mastronardi, C. A., Licinio, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptin+treatment%3A+facts+and+expectations%5BTitle%5D)+AND+%22Metabolism%22%5BJournal%5D)">Leptin treatment: facts and expectations.</a> Metabolism. 64 (1), 146-156 (2015).</li>
</ol>

<table><tbody><tr><td>Soluzione alcolica di clorexidina digluconato allo 0,5%</td><td>Rioqu&iacute; Mica</td><td>SKU 1068253</td><td></td></tr><tr><td>Ago calibro 16</td><td>Becton Dickinson</td><td>SKU 305198</td><td></td></tr><tr><td>Soluzione di digluconato di clorexidina non alcolica al 2,0%</td><td>Rioqu&iacute; Mica</td><td>SKU 1113045</td><td></td></tr><tr><td>Siringa da 3 mL</td><td>Becton Dickinson</td><td>SKU 309657</td><td></td></tr><tr><td>Strisce Accu-Check e ndash; Performa</td><td>Roche</td><td>EAN 4015630981960</td><td></td></tr><tr><td>Alcian Blue PAS Kit</td><td>EasyPath</td><td>SKU EP-11-20019</td><td></td></tr><tr><td>Albumina Sirica Bovina (BSA)</td><td>Sigma-Aldrich</td><td>A4612</td><td></td></tr><tr><td>Software CellSens</td><td>Olympus</td><td>CELLSENS</td><td></td></tr><tr><td>Citrato Buffer pH 6.0: Acido citrico monoidrato + Idrossido di sodio</td><td>Sigma-Aldrich</td><td>C1909 + 28-3010</td><td></td></tr><tr><td>Anticorpo della Caspasi-3 tagliata</td><td>Tecnologia di segnalazione cellulare</td><td>9661</td><td></td></tr><tr><td>Analizzatore Cobas C 111</td><td>Roche</td><td>4528778001</td><td></td></tr><tr><td>Creatinina K</td><td>Labtest</td><td>96-300</td><td></td></tr><tr><td>EnVision FLEX DAB+ Chromogen</td><td>Agilent-Dako</td><td>K3468</td><td></td></tr><tr><td>Reagente di rilevamento EnVision FLEX /HRP</td><td>Agilent-Dako</td><td>SM802</td><td></td></tr><tr><td>Reagente Envision FLEX che blocca la perossidasi</td><td>Agilent-Dako</td><td>SM801</td><td></td></tr><tr><td>Buffer Substrato EnVision FLEX</td><td>Agilent-Dako</td><td>SM803</td><td></td></tr><tr><td>EnVision FLEX era buffer 20x</td><td>Agilent-Dako</td><td>K800721-2</td><td></td></tr><tr><td>Eosina</td><td>Agilent-Dako</td><td>CS701</td><td></td></tr><tr><td>Lettore di microplacche GloMax Discover</td><td>Promega</td><td>GM3000</td><td></td></tr><tr><td>Glucometro Accu-Check - Performa</td><td>Roche</td><td>EAN 4015630980512</td><td></td></tr><tr><td>Software GraphPad Prism 8 (software statistico)</td><td>GraphPad di Dotmatics</td><td>-</td><td></td></tr><tr><td>100% v/v Isoforina (Isoflurano)</td><td>Crist&aacute; Lia</td><td>-</td><td></td></tr><tr><td>Ematossilina</td><td>Agilent-Dako</td><td>CS700</td><td></td></tr><tr><td>Microscopio della luce IX51</td><td>Olympus</td><td>IX51</td><td></td></tr><tr><td>Gabbia metabolica per topo singolo</td><td>Tecniplast</td><td>3600M021</td><td></td></tr><tr><td>Mezzo di montaggio - Entellan</td><td>Sigma-Aldrich</td><td>100869</td><td></td></tr><tr><td>Kit ELISA per albumina topo</td><td>Abcam</td><td>Ab207620</td><td></td></tr><tr><td>Ceppo di topo: BTBR. Cg-Lepob/WiscJ</td><td>JAX Laboratories</td><td>4824</td><td></td></tr><tr><td>Periodic Acid Schiff (PAS) Kit</td><td>EasyPath</td><td>EP-11-20014</td><td></td></tr><tr><td>RM255 Microtomo</td><td>Leica Biosystems</td><td>14050237960</td><td></td></tr><tr><td>Processore di tessuti TP1020</td><td>Leica Biosystems</td><td>TP-1020</td><td></td></tr><tr><td>Tramadolo 50 mg/mL</td><td>Gr&uuml; Nenthal</td><td>4022444</td><td></td></tr><tr><td>Anticorpo WT-1</td><td>Santa Cruz</td><td>SC-192</td><td></td></tr></tbody></table>

innesti adiposi bianchi e marroni: un approccio per correggere i deficit riproduttivi, metabolici e renali nei topi obesi black and tan brachyury (btbr)

White and Brown Adipose Grafts: An Approach to Correct Reproductive, Metabolic, and Renal Deficits in Black and Tan Brachyury (BTBR) Obese Mice

Innesti adiposi bianchi e marroni: un approccio per correggere i deficit riproduttivi, metabolici e renali nei topi obesi Black and Tan Brachyury (BTBR)

Transplantation of white adipose tissue (WAT) and brown adipose tissue (BAT) has emerged as a promising therapeutic strategy for reversing metabolic ...

white-brown-adipose-grafts-an-approach-to-correct-reproductive

Research

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JoVE Journal

Biology

Biologia

In questo video, dimostriamo la procedura chirurgica per isolare il tessuto adiposo bianco sottocutaneo e il tessuto adiposo bruno interscapolare da un topo appena nato.

Raccolta del deposito adiposo bianco e marrone dal cucciolo di topo: una procedura chirurgica per raccogliere il tessuto adiposo bianco e il tessuto adiposo bruno dal cucciolo di topo

Primarie preadipociti bianchi isolati dal tessuto adiposo bianco nei topi possono essere differenziate in cellule del beige / brite. Presentato qui è un sistema affidabile modello cellulare per studiare la regolazione molecolare della "doratura" di grasso bianco.

Isolamento e la differenziazione di cellule stromali vascolari alle celle Beige / Brite

Questo articolo descrive un metodo di trapianto per innestare cellule epiteliali mammarie del donatore nel cuscinetto di grasso bianco interscapular degli animali riceventi. Questo metodo può essere utilizzato per esaminare gli effetti dell'ospite e/o del donatore sullo sviluppo dell'epitelio mammario ed elimina la necessità di pre-clearing, estendendo così l'utilità di questa tecnica.

Rat mammary Epithelial Cell Transplantation nel Pad Grasso Bianco Interscapular

Questo rapporto descrive un protocollo per l'isolamento simultaneo dei preadipociti primari marroni e bianchi dai topi appena nati. Le cellule isolate possono essere coltivate in coltura e indotte a differenziarsi in adipociti bianchi e marroni completamente maturi. Il metodo consente la caratterizzazione genetica, molecolare e funzionale delle cellule adipose primarie in coltura.

Isolamento e differenziazione dei preadipociti bianchi e marroni primari dai topi appena nati

Imaging funzionale e quantificazione di depositi adiposi termogenici nei topi utilizzando un approccio basato sull'imaging micro-PET/MR.

Visualizzazione e quantificazione dei tessuti adiposi marroni e beige nei topi utilizzando [18F]FDG Micro-PET/MR Imaging

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

Qui, presentiamo un protocollo dettagliato che delinea l'uso di frammenti microvascolari isolati da roditori o tessuto adiposo umano come approccio diretto per progettare tessuto adiposo beige funzionale e vascolarizzato.

Coltura tridimensionale di tessuto adiposo termogenico vascolarizzato da frammenti microvascolari

Questo protocollo presenta l'uso di CRISPR SunTag-p65-HSF1 (SPH) negli adipociti (AdipoSPH) come strategia alternativa al virus adeno-associato (AAV) per studiare la biologia del grasso beige. L'iniezione in vivo di sgRNA portatori di AAV mirati al gene endogeno Prdm16 è sufficiente per indurre lo sviluppo di grasso beige e migliorare il programma genico termogenico.

Studio della biologia e del metabolismo dei grassi beige utilizzando il sistema di attivazione CRISPR SunTag-p65-HSF1

In questo protocollo, descriviamo le procedure tecniche per generare topi knockout specifici per il tessuto adiposo bruno (BAT) sfruttando un sistema combinato di Cre-LoxP, CRISPR-Cas9 e RNA a guida singola (sgRNA) del virus adeno-associato (AAV). I passaggi descritti includono la progettazione degli sgRNA, la preparazione delle particelle AAV-sgRNA e la microiniezione di AAV nei lobi BAT.

Generazione di topi knockout specifici per il grasso bruno utilizzando un sistema combinato di RNA a guida singola Cre-LoxP, CRISPR-Cas9 e virus adeno-associato

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Scopri il trapianto combinato di tessuto adiposo bianco e bruno per correggere disfunzioni metaboliche e riproduttive nei topi obesi BTBR. Questo protocollo standardizzato migliora la fertilità, il metabolismo e la funzione renale.

Innesti adiposi bianchi e marroni: un approccio per correggere i deficit riproduttivi, metabolici e renali nei topi obesi Black and Tan Brachyury (BTBR) (Video) | JoVE Journal

Innesti adiposi bianchi e marroni: un approccio per correggere i deficit riproduttivi, metabolici e renali nei topi obesi Black and Tan Brachyury (BTBR)

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