• While co-culturing both vascular endothelial and smooth musclecells in a filtering culture cup, the environmental stimulus can beprovided with them.
• Environmental stimulus similar to live organs is given tovarious kinds of cells, and then respective gene, and proteinlinked with them are analyzed at molecular scale routinely.
• Saves the culture media and shortens the time of collection andrefinery of target samples, as only a little quantity of culturesolution is circulated.
• Filtering culture cups are available on the market, andpossible to co-culture previously, which allows you to obtain thegood repeatability by handling and seeding cellseasily.

1. Analysis of gene expression (Human vascular endothelial cells)in all respects caused by the combined stimulus of flow, Oxygenconcentration gradient and Low-density lipoprotein (LDL)

2. Analysis of anti-inflammation's symptoms caused by flow stress(Human vascular endothelial cells, vascular endothelial cells oftranscription factors knockout mouse)

[ Effect on Endothelial Cell Gene Expression of ShearStress, Oxygen Concentration, and Low-Density Lipoprotein asStudied by a Novel Flow Cell Culture System ]
Free Radic Biol Med. 2004 Sep 1;37(5):682-94.

• The culture cell of this system is divided into upper and lowercells by the partition of a permeable PET membrane.
• A cell culture insert for 6-well plates manufactured by BD isused as the filtering culture cup.
• The cells of membrane can be exposed with the physical power offlow generated by the circulation of 10cc of culture solution.
• As Nitrogen gas can be replaced with the air of the culturecell, an oxygen concentration gradient is performed by thecirculation of culture solution having a very low dissolved oxygenconcentration--its partial pressure of oxygen is about 2 to 5 %.
• An observation window is provided for an inverted or a co-focusmicroscope.
• A diaphragm pump with the minimum differential pressuremaintains the culture solutions of upper and lower cupwell-balanced for a long time--the operation is available overnightwithout an operator present.
• Possible of monitoring the partial pressure of dissolvedoxygen.
• The drug injection or the sampling after the drug injection isavailable via a drug sampling port.
• All contaminated parts can be autoclaved.

1. Culture cell cassette
2. Solution tank
3. Diaphragm Pump

• Low shear stress (about 0.2 to 2 dynes/cm2)
• Seeding cells in the filtering culture cup
• Long keeping the oxygen low concentration gradient

• High shear stress (about 5 to 15 dynes/cm²)
• Seeding cells at outside of filter is to be loaded high shearstress

The air in the culture cell was replaced with Nitrogen gas so asnot to generate any gases inside while the culture solutioncontaining serum was circulated--for each 15ml of upper and lowerchambers.

The measuring points of DO are shown in the left sketch. Hypoxiccondition (2-5%) appeared in some 10 minutes.

Pump flow rate: 3-30 cc/min
Filter pore size: 0.4 micro meter dia.
N2 flow rate:50 cc/min
Measuring point: (1)

 The maximum descent velocity of lowconcentration of Oxygen is 15cc/min.

Pump flow rate: 3, 9 cc/min
Filter pore size: 0.4, 3.0 micro meter dia.
N2 flow rate:50 cc/min
Measuring point: (1)

In case that the large flow rate is set, there is atendency that the larger pore size of filter, the lesser effectiveexchange rate of DO.

Pump flow rate: 3 cc/min
Filter pore size: 0.4, micro meter dia.
N2 flow rate:50 cc/min
Measuring point: (1) to (4)

The performance of an oxygen concentration gradientin about 30 min is confirmed.

The following accessories are optional at extra charge.

1. CO2 incubator
2. Gas cylinders
3. When the low oxygen concentration is desired in operation, anoptional incubator and the gas exchange system are necessary,although they are not shown in the photos.
4. The system does NOT include vascular wall cells. Please contactus regarding the method of culture.