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Temperature-responsive plate RepCell

Cell culture plate that detach cells by temperature reduction
 

Temperature-responsive plate

 
RepCell®


 

Key features of RepCell®
♦Temperature responsive polymer is fixed to the surface of 96 multi-well plate at nano-thickness level.
♦Cells are cultured at 37°C and collectable approximately 30min after reducing temperature down to 20°C.

Advantages of RepCell®
Collection of cells by mere reduction of temperature
♦It completely abandoned the usage of trypsin when collecting cells since cells deatch by just lowering the temperature. Saves time for laborious pipetting procedure. Our internal report clarified that total labor spent on these procedures were cut down to 1/5.

♦Suitable for limited dilution methodology for cell cloning.



No damage to the cells
♦Trypsin is well known for its deterioration to the functions of cells. Maintenance of their activities is possible when RepCell is used for cell culture.

♦Suitable for culturing fragile cells such as stem cells or neurons.

♦It can also be applied to the study of extra cellular matrix or surface antigens.

♦Cell therapeutics can also be produced without enzymatic contamination.

Save cost on pipett chips
♦Approximately US$10 can be saved if conventional methodology is replaced with RepCell.

  Conventional method RepCell
Cost of plate (1 piece) US$4 US$22
Sterlized chips@US$0.1
(Replaced when sucking media and PBS)		 96wellsx4timesxUS$0.1=$38 96xoncexUS$0.1=$9.6
PBS50ml($1.6)·Trypsin100ml($2) US$3.6 Not required
Total Cost spent for cell cloning per plate US$45.6 US$31.6

 

Suitable cell type for RepCell®
♦RepCell is suitable culturing cells shown in the following diagram
 


 

INSTRUCTION
RepCell
Culturing cells
♦The surface becomes hydrophobic above 32°C therefore cells must be cultured above this point
(we recommend to culture cells at37°C)

♦Media change can be carried out as if using normal plate without polymer.
(Attachment time of the cells may vary depending on the type of the cell. )

Detachment of cells
♦The surface of the plate becomes hydrophilic under 32°C. Cells are easier to detach under this temperature.

♦When collecting cells, lower the temperature down to approximately 20°C and wait about 30 minutes. Depending on the type of the cells cultured, they detach automatically.
(Same results can be achieved by leaving the plate in the refrigerator set at 4°C for 30 minutes.)

♦The cells that did not come off automatically can be detached by agitating them by pipet with media.
(Some cells may not detach completely. The results may vary by the type of the cells.)

♦Detachment of cells are easier when the temperature is lowered before cells become confluent.

♦The product is intended for one time use only. Please do not re-use the product.

Wells/Bottom shape 96(6f)/Flat
Capacity per well 0.3 ml
Size 86mm(L) X 128mm(W)
Material Polystyrene (with lid)
Area per well 0.32 cm2

Temperature-responsive plate

RepCell(R)


 		  
Microscopic view of cell culture/detachment of using "RepCell"

From the data below, it can be concluded that...
1) Cells are detaching naturally by mere reduction of temperature
2) The damage given to the cells due to trypsin is so severe that it took 5 times more time than that of temeperature reduction till they re-attached to the surface of the dish.

(1)Detachment of cells by reduction of temperature.

 
Normal culture(37°C) 30min post-cool(20°C) Removal after
slight agitation

A549
A549
A549

Swiss 3T3
Swiss 3T3
Swiss 3T3

NRK
NRK
NRK

Mesenchymal stem cells
Mesenchymal stem cells
Mesenchymal stem cells

Peritoneal macrophage
(Mice)
Peritoneal macrophage
(Mice)
Peritoneal macrophage
(Mice)

(2)Comparison of re-attachment kinetics for harvested A549 cells:Trypsin mediated vs Temperature harvest.

 
10min 60min 120min 300min

Trypsin mediated
Trypsin mediated
Trypsin mediated
Trypsin mediated

Temperature harvest
Temperature harvest
Temperature harvest
Temperature harvest
(3)Comparison of re-attachment kinetics for harvested A549 cells:Trypsin mediated vs Temperature harvest.

 
Cell culture plate that detach cells by temperature reduction

Temperature-responsive plateRepCell(R)




 		  
Q. What kind of cells are you targeting?
A. You can culture almost all the endothelial cells such as NIH/3T3, Swiss/3T3 or 10T1/2. As far as epidermal cells are concerned, you can culture them as long as its adherence is not too strong. Please note that some cells whose adherence is strong may not come off as expected.
Q. Isn't the price too expensive?
A. The advantage of using our product when compared to non-coated products are as follows:
1) If you detach cells by using trypsin, you would require to change pipett chip to new one so as to alleviate contamination when sucking media, and sucking PBS for the first and second time. This renders its total cost to be more expensive than the price of RepCell.

2) Conventional methodology would require 8 times more of pipetting (sucking media, add PBS for the first time, suck PBS, add PBS for the second time, suck PBS, add trypsin and agitation) than RepCell which only requires reduction of the temperature when collecting cells. This leads to the reduction in time used for cloning cells. You can allocate your precious time for more productive work such as writing publications. We have an internal report that suggests the total time spent on the cell cloning procedures were reduced down to 1/5.

3) Because the total procedure could be cut down to 1/5, you can avoid the risk of cross-contamination derived from fatigue, which will eventually improve the accuracy of cell-cloning procedure.

4) Since you do not need to use trypsin to collect cells, the activity of the cells are maintained.
 
Q. Does the collected grouped cells become single/independent?
A. The polymer coated surface allows collection of grouped cells without damaging its cell-cell junctions. Therefore the collected cells do not automatically become independent or single. However, those cells that has just been divided sometimes become single/independent by pipett agitation. This occurs because cell-cell junction is not so strong if the cells have just been divided.
Q. Do you need temperature control unit which can be lowered down to 20°C?
A. You can achieve the same result by leaving the plate in refrigerator set at 4°C for 30 minutes.

 

 

 

For more information :: Bioxys and Gentaur BVBA :: Av. de l'Armée 68 B4 :: BE-1040 BELGIUM

Email: info@gentaur.com
 

International 

+32 (0) 16 58 90 45

+32 (0) 16 50 90 45

France

01 43 25 01 50

01 43 25 01 60

Italy

02 36 00 65 93

02 36 00 65 94

Germany

0241 6085 13140

0241 6085 33033
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Last modified: feb-07