Q1. What cell lines can be grown on the BelloCell bioreactor?

All commonly-used attachment cells have been grown on the BelloCell bioreactor including VERO, CHO, MDCK and HEK 293 cells.

Q2. What is BelloCell advantage superior to other competitor’s products?

The mechanism of BelloCell enables the fast, complete diffusion of nutrients and oxygen in the medium to the matrix, on which cell resides. All ingredients in the medium are mixed well while the CO2, which is produced by glucose hydrolysis, can be expelled efficiently during the run.

Q3. Is BelloCell patented?

BelloCell systems are well patented and the patent covers US, Europe, Japan, China and Taiwan.

Q4. How many matrices are in BelloCell-500 family?

There are regularly 865 pieces of BioNOC II in each BelloCell bottle. The deviation is less than 5%.

Q5. How can I detach the cells growing on the BioNOC II carrier in the BelloCell?

There are several ways to solve the issue:

<i> To use enzyme other than trypsin, for example, Accutase. Some other enzyme but not trypsin-based may also be helpful.  

<ii> To disrupt the cells from the carriers through freeze-and-thaw, with lysis buffer, or other methods. 

Q6. How many pieces of carriers in BelloCell are required for cell counting? Can I put the carriers i

It is suggested to do triple duplicate and then stain with CVD and then multiply averaged calculated cell number by 865 (piece/ BelloCell bottle). The result would be the total cell numbers. Before trypsinization, check the reference to make sure that cells can be detached. Usually it takes more efforts to detach the bigger cells.

Q7. What is the cell productivity with respect to medium volume (# of cell/ ml of medium) of BelloCel

Without changing the gene of cells, it is very unlikely to promote the cell propagation while limiting critical nutrients. With the same quantity of medium in a certain period of time, BelloCell’s maximum output may not be different from those of other commercial products. However, BelloCell does present higher cell density due to the cell growth pattern.

Q8. What is the maximum productivity of one BelloCell? Is it easy to harvest the cells?

Generally speaking, 5 × 109/ml of cells (depending on cell types) is an acceptable output in one BelloCell. The harvest of cells from BelloCell is quite easy. Follow the SOP provided by CESCO and more than 90% of recovery is not rare.

Q9. Can I use BelloStage 3000 with BelloCell-500E for suspension culturing of the tumor cells without

BelloCell only works for adherent cell culture. However, if the specific suspension cells could attach in the BelloCell system, the secreted protein can be harvested for long periods with much lower cost than commercial suspension culture system.

Q10. BelloFeeder 1300 is only for 3 tubing sets and BelloStage 3000 is for 4 Bello bottles at a time-

BelloFeeder 1300 was originally designed in a standard 6 ft3 CO2 incubator.  Once three BelloCell bottles are operated, there will required 3x 2 L glass vessel, 3x tubing complete sets, 1x BelloFeeder-1300 and 1x BelloStage-3000 which will occupy the entire space inside the incubator. If customers get larger incubation space and could run 4x BelloCell bottles, we have another BelloFeeder-1100 with only single pump head.

Q11. Can I use BelloCell-500A or BellCell-500AP in cell culture for production of secreted product?

Yes. BelloCell-500A or BelloCell-500AP is suitable for production of both secreted and non-secreted products. Since these two types of bottles are specifically designed to enhance the cell attachment efficiency, we suggest to use them with the up-side-down seeding protocol if your cell line has difficulties to adhere the carriers.

Q12. What if the level of the ingress protection of BelloStage?

The main console of the BelloStage meets IP54.

Q13. Can the parameters we set up in BelloCell-500 be applied in TideCell-002? If so, is this positiv

BelloCell-500 is used mostly in the lab for research and tissue engineering whereas TideCell-002 can be applied in both academy and small pilot tests. The final cell densities in both systems are quite related proportionally. However, the productivity and cell growth rate depend on several factors.  

Since BelloCell-500 doesn’t have pH control and perfusion culture mode, the cell growth usually is slower than in TideCell-002, under a manual operation. With respect to the linearity of scale-up, TideCell-002 is better than BelloCell-500. The parameter setting in BelloCell-500 could be used in TideCell-002, but some modifications are still necessary.  Compared with other commercially-available systems, however, very few parameters are required to input in the TideCell system. Therefore optimization is quite straight -forward.  

Q14. We usually coat our cell culture surfaces with Matrigel (BD) for HEK cell line. Would that be OK

The surface of the BioNOC II carriers in the BelloCell is negative charged and should be OK for the coating of Matrigel if it could be coated on petri dish or T-flask (those are all negative charge surface). Some sources of HEK293, for example, are difficult to attach, and therefore we would make coating or use BelloCell-500A to enhance the cell attachment efficiency.

Q15. Documentation related to the validation on the BelloCell system?

CESCO Bioengineering is an ISO 9001:2008 certificated company. The quality documents for BelloCell include the Certificate of Qualification (CoQ) and Certificate of Compliance (CoC). The BelloStage includes the CE certificate and Certificate of Qualification (CoQ).  

Q16. Can the standard BelloCell bottle be frozen?

Yes. The bottle could be placed in -80 °C freezer directly for free-and-thaw process to extract cellular components.

Q17. How can I maintain VERO cells in serum free medium for 10 days in BelloCell?

Once the nutrients of medium are consumed, you need to refresh culture medium. To maintain 10 days without exchanging culture medium, under high cell density, is very difficult unless the culture temperature can be reduced to below 10 C. Or supplement such as glucose and glutamine concentrate should be added to maintain the cell viability. Be sure to control pH within the proper range (7.0~7.4) by adding concentrated NaHCO3. However, the osmolarity will increase and affect cell viability if too much sodium bicarbonate is added.

Q18. How can we conclude that the cells have reached maximum density and may not further grow stage?

It would be required to run the cell growth test and monitor the cell density every other day until the growth of cells reaches the plateau. However, for the cell line, it should reach around 1.5~2x109 cells per BelloCell bottle from our past experience.

Q19. A customer is using BelloCell-500 A. A question is that if the infection procedure is similar to

It depends on how the viruses infect cells. If the viruses could effectively infect the cells, he could do infection as the regular way as cell seeding.

Q20. For virus infection, how much volume do you suggest the users use? 100ml, similar to cell seedi

Usually 100~120 ml is sufficient.

Q21. For trypsinization, do you suggest the user to place bottle upside down similar to seeding to ma

Yes. For the cell harvest, it is better to use BelloCell-500A since it requires less trypsin to detach cells, less wash buffer, and has higher cell recovery rate.

Q21. For trypsinization, do you suggest the user to place bottle upside down similar to seeding to ma

Yes. For the cell harvest, it is better to use BelloCell-500A since it requires less trypsin to detach cells, less wash buffer, and has higher cell recovery rate.

QWhen does the AF-1200 monitor feed and harvest weights?

 AF-1200 monitors the summation  of "NET WEIGHT" of feeding reservior and harvest reservior only between the (n)th and (n+1)th medium exchange cycle.

QWhat is the control logic of AF-1200 feed and harvest pump?

 If total weigh of net weight  > upper protection weight, harvest pump will act for 30 sec and stop. (it is called total weight recorvery procedure).

AF-1200 will detect total weight again to make sure total weight <= upper protection weight. If not, harvest pump will act another 30 sec and stop, repeating until total weight <= upper protection weight.

QHow to calculate pump startup frequency

 If the medium exchange parameters of N step is 250L/Day ; 24 hr, 2L/cycle.  

The interval between each cycle of medium exchange is (24*60)/(250*24/24/2) = 11min , decimal is eliminated by PLC.