Corning® BioCoatTM 355057 • Falcon Entero-STIM • • Entero-STIM acid Life Sciences Butyric © 2013 Corning Incorporated 2 Corning BioCoat 355057 Corning BioCoat – – 24 24 ENTERO-STIM MITO+ m 24 353504 ×1 ×1 - 100 mL - 400 mL -1 355057 Life Sciences © 2013 Corning Incorporated 3 250 mL 2 355058 ENTERO-STIM 250 mL 2 MITO+ 500mL ENTERO-STIM 355357 250 mL MITO+ 355006 5L Corning BioCoat 354472 1.0 m 24 Life Sciences 354474 2 24 /6 24 /24 24 © 2013 Corning Incorporated 4 • 353502/353504 – 353507 353502 • 353507 – 353502/353504 • • • • • • * Falcon Life Sciences © 2013 Corning Incorporated 5 24hrs 48hrs *3 Life Sciences © 2013 Corning Incorporated 6 • MITO+ 70 • MITO+ 100 l 500 l 400 l 125ml 500mL MITO+ 100 l MITO+ 400 l 125ml MITO+ 500mL 2 8 Life Sciences 21 © 2013 Corning Incorporated 7 • 250,000cells/cm2 • Life Sciences © 2013 Corning Incorporated 8 24 1. Caco-2 2. /EDTA Caco- 2 3. 4x105 /EDTA MITO+ 4x105 /mL 4. 37 /ml MITO+ (37 ) 1m 500 l 5. • 6. 37 200,000 5%CO2 100% 500µl / 24 1mL/ Life Sciences © 2013 Corning Incorporated 9 1. – – 2. 37 MITO+ 1mL 3. 4. Life Sciences 500 l 37 5% CO2 48 100% © 2013 Corning Incorporated 10 1. 2. 3. PTEER Life Sciences © 2013 Corning Incorporated 11 1. – Life Sciences © 2013 Corning Incorporated 12 2. – Life Sciences © 2013 Corning Incorporated 13 3. – 3 * Life Sciences © 2013 Corning Incorporated 14 3. – 3 * Life Sciences © 2013 Corning Incorporated 15 • 48hrs 24hrs 24hrs 24hrs New and better protocols for a short-term Caco-2 cell culture system. Yamashita S, Konishi K, Yamazaki Y, Taki Y, Sakane T, Sezaki H, Furuyama Y. J Pharm Sci. 91, pp669 2002 Life Sciences © 2013 Corning Incorporated 16 • Trans Epithelial Electrical Resistance TEER) – • Paracellular Markers – Lucifer yellow - mw 487 – Mannitol - mw 182 – Polyethylene Glycol (PEG) (400 - 4000) - mw 400 - 4000 • Visualization * – – – – * 3 2 Life Sciences © 2013 Corning Incorporated 17 Transepithelial Electrical Resistance (TEER)TEER TEER = (Rm-Rb) x A TEER Rm Rb A transepithelial electrical resistance [Wcm2] [W] [W] [cm2] Note: TEER * Life Sciences © 2013 Corning Incorporated 18 Lucifer yellow • Lucifer Yellow Flux (% LY Flux) % LY Flux = 100 x ((LYBL x VolBL) / (LYAP x VolAP)) LYBL VolAP LYAP VolBL Note: Lucifer Yellow Lucifer yellow [cm3] Lucifer yellow M] [cm3] 0.3 to 2% M] 2% • Drug Transport Assay for Falcon® 24-Multiwell Insert Systems Life Sciences © 2013 Corning Incorporated 19 • Drug Transport Assay for Falcon® 24-Multiwell Insert Systems Transport Buffer HBSS (with Ca++, Mg++) + 10 mM HEPES, pH 7.0 – 7.4 Ca++ /Mg++ pH pH apical pH 6.4, basal pH 7.4 Temperature 37 transcellular transport active transport Life Sciences © 2013 Corning Incorporated 20 03-5386-1996 [email protected] Life Sciences © 2013 Corning Incorporated 21
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