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Corning® BioCoatTM
355057
Life Sciences © 2013 Corning Incorporated 2
• Falcon®
Entero-STIM
•
• Entero-STIM Butyric acid
Life Sciences © 2013 Corning Incorporated 3
Corning® BioCoatTM
355057
Corning BioCoat m
– 24 24 ×1– 24 353504 ×1
- 100 mLENTERO-STIM - 400 mLMITO+ - 1
355057
Life Sciences © 2013 Corning Incorporated 4
355058
250 mL 2
ENTERO-STIM 250 mL 2
MITO+ 500mL
ENTERO-STIM 355357 250 mL 2
MITO+ 355006 5L
Corning BioCoat
1.0 m
35447224 /6
24 35447424 /24
24
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•
• *
• Falcon
• 353502 6 /353504 24/
•
•
•
353502
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24hrs 48hrs
*3
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MITO+
100 l 400 l
125ml 500mL
• MITO+ 70
• 500 l
MITO+ 2 8 21
125mlMITO+
100 l
500mLMITO+
400 l
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•250,000cells/cm2
•
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24
1. Caco-2
2. /EDTA Caco-2
3. /EDTA 37MITO+
4x105 /ml
4. MITO+(37 ) 1m
5.500µl
• 200,000 /
6. 37 5%CO2 100% 24
4x105 /mL
500 l
1mL/
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1.
––
2.37 MITO+
1mL
3. 500 l
4. 37 5% CO2 100% 48
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1.2.3.
P-TEER
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1.
–
2.
–
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3.
– 3
*
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3.
–3
*
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•
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 *– –
– –
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Transepithelial Electrical Resistance (TEER)TEER
TEER = (Rm-Rb) x ATEER transepithelial electrical resistance [Wcm2]Rm [W]Rb [W]A [cm2]
Note: TEER
*
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Lucifer yellow
• Lucifer Yellow Flux (% LY Flux)
% LY Flux = 100 x ((LYBL x VolBL) / (LYAP x VolAP))LYBL Lucifer yellow M]VolBL [cm3]LYAP Lucifer yellow M]VolAP [cm3]
Note: Lucifer Yellow 0.3 to 2% 2%
•Drug Transport Assay for Falcon® 24-Multiwell Insert Systems
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•Drug Transport Assay for Falcon® 24-Multiwell Insert Systems
Transport Buffer HBSS (with Ca++, Mg++) + 10 mM HEPES, pH 7.0 – 7.4Ca++ /Mg++
pHpH apical pH 6.4, basal pH 7.4
Temperature 37 transcellular transport active transport
