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Corning® BioCoatTM

355057

Life Sciences © 2013 Corning Incorporated 2

• Falcon®

Entero-STIM

•

• Entero-STIM Butyric acid

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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

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• Trans Epithelial Electrical Resistance TEER)–

• Paracellular Markers– Lucifer yellow - mw 487 – Mannitol - mw 182– Polyethylene Glycol (PEG) (400 - 4000) - mw 400 - 4000

• Visualization *– –

– –

* 32

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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

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03-3586-1268ScientificSupportJP@corning.com