Navigation Links
CHO Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor


Introduction
This Application Report is one in a series documenting simple protocols for culturing cells in a BioFlo 110 benchtop fermentor/bioreactor system. The modular BioFlo 110 is offered with an extensive choice of vessels, impellers and control modules to enable growth of a wide variety of cultures, including mammalian, insect, plant, yeast and bacterial cells. Complete kits are also available. This report concentrates on achieving high-density CHO cell cultures using NBS Advanced Cell Culture Kit.

For this application, researchers in NBS cell culture laboratory used a:
BioFlo 110 Advanced Cell Culture Kit with a magnetic drive and a 3.0L total volume water-jacketed vessel, (nominal working volume 2.2L.) (NBS Catalog # M1273-1212)
Suspension Cell Spin Filter with a 12-14 micron screen (NBS Catalog # M1273-3202) for continuous perfusion culture
BioCommand Plus Supervisory Software (NBS Catalog # M1291-0000) to control the feeding schedule and collect data
NucleoCounter Automated Cell Counting System (NBS Catalog # M1293-0000) to count viable cells.


The Bioreactor
All BioFlo 110 Advanced Cell Culture Kits are configured with six control modules, a complete vessel assembly including a pitched blade agitation impeller, foam/level sensors, as well as dissolved oxygen and pH probes (Mettler Toledo), three liquid addition bottles, cables, tubing, clamps and other related items. Comes with choice of direct drive or magnetic-drive agitation systems, as well choice of water-jacketed or heat-blanketed vessel options.

For this application we substituted an optional NBS spin filter impeller for the standard pitched-blade impeller. The spin filter impeller is very useful for a continuous perfusion process because it is a simple device that lends itself to media exchange and can be easily scaled up. With most commonly available spin filters, screens rapidly become clogged when cells reach high cell density during perfusion. NBS, however, has designed spin filters with three different screen sizes that are suitable for most cell types. Feel free to contact NBS for screen size suggestions.




Control System
Listed below are six control modules included in an Advanced Cell Culture Kit, which were also used for this application:


Materials and Methods

Overview

Sterilize the vessel with phosphate buffer solution (PBS) for 60 minutes.
Remove the PBS from the vessel.
Add 1.5 L of media into the vessel.
Key in control setpoints.
Inoculate 500 mL
Use BioCommand software to automatically calculate and control glucose addition throughout the run.

Medium
EX-CELL 325 PF CHO from JRH.

Cell Line: CHO/B4C7

PCU: Select Cell Culture Application
On the initial set-up screen, the USE selector button will allow you to change the default Fermentation application to Cell Culture application. For further details refer to the Operation Section in the BioFlo 110 manual.

Control Setpoints
Setpoints were keyed into the controller prior to inoculation. The vessel was allowed to equilibrate prior to inoculation. The DO loops Current Value remained higher than setpoint until the culture was introduced.
Temperature . . . . . . . . . . . . . . . .37C
pH . . . . . . . . . . . . . . . . . . . . . . . .7.2
DO . . . . . . . . . . . . . . . . . . . . . . . .50%
Agitation . . . . . . . . . . . . . . . . . . .70-130 rpm
O2 (Gases) . . . . . . . . . . . . . . . . . .4 Gas

PH Control
The pH probe was calibrated prior to the autoclave cycle (refer to BioFlo 110 Instruction Manual). CO2 gas and a liquid base were used to maintain the pH setpoint. pH control parameters were:
Base . . . . . . . . .. . .Sodium bicarbonate, 8% solution
Pump . . . . . . .. . . .Pump 4 of the 4-Pump Module
Transfer tubing . . .Narrow bore silicone tubing, as supplied (116" ID & 316" OD - NBS Catalog # P0740-2396)
Vessel inlet . . . . .. .Triport adapter in the vessel headplate

PCU setup: (pH loop)
Pump 4 plugged into "BASE" Power Controller Outlet
pH Control Selections:
Auto mode
Multiplier = 100% until base feed is started, after inoculation. Then, 25%
Multiplier = 25% after inoculation
Dead-band = 0.1
PID values: factory defaults

Dissolved Oxygen (DO) Control
The DO probe was calibrated after the autoclave cycle (refer to BioFlo 110 Instruction Manual). DO control was set to Auto mode, which automatically defaults to 4-Gas Mode after the Gas Control (O2 loop) selection is made. We also changed the P and I value of the DO loop to meet the culture demands, as shown on the next page.

PCU Setup: (DO loop)
DO Control Selections: Auto (4 Gas mode)
PID values: P = 2.00 and I = 0.67

Gas Control (O2 loop)
The control was set to 4-Gas mode, to automatically maintain DO and pH setpoints, using 3 gases (Air, O2, N2) to control the DO setpoint and CO2 to control pH setpoint.

PCU Setup: (O2 loop)
O2 Control Selections: 4 Gas mode

Continuous Feed
All pumps were calibrated using the standard tubing supplied to keep track of the liquid quantities entering and exiting the vessel.

Samples were taken several times a day for off-line measurement of glucose and cell density.

PCU setup: (Pump 1 & Pump 2 loop)
Pump 1 . . . . . . . . . . .. .plugged into "Pump A" Power Controller outlet
Pump 2 . . . . . . . . . .. . .plugged into "Pump B" Power Controller outlet
Pump 1 (Nutrient) . .. ..Vessel inlet - Triport adapter in the vessel headplate
Pump 2 (Harvest) . . . .Vessel outlet - 12mm port using the dip tube
Pump A Selection . .. . .Manual mode (automatically adds liquid to the vessel with the BioCommand control program)
Pump B Selection . .. . .Wet On (removes liquid from the vessel using the level probe as a trigger to turn the pump on)

Control Program
For this study, we used NBS BioCommand Plus software to control nutrient addition. For the nutrient control program, we calculated the approximate time frame profile to automatically add fresh media using pump 1.




'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Big Blue Rodent Cell Cultures for Mutagenesis Studies
2. Simple Isolation of RNA from Tissue and Cultured Cells
3. The QIAGEN Guide to Animal Cell Culture
4. The QIAGEN Guide to Animal Cell Culture
5. The QIAGEN Guide to Animal Cell Culture
6. The QIAGEN Guide to Animal Cell Culture
7. The QIAGEN Guide to Animal Cell Culture
8. The QIAGEN Guide to Animal Cell Culture
9. RNAi in Non-Mammalian Cultured Cells
10. Total RNA Isolation: 100 l Viral Sample From Cultured-Cell Supernatant Containing Virus
11. Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: CHO Cell Culture Application Using BioFlo Benchtop Bioreactor

(Date:4/23/2015)... Six startups from Russia traveled ... as part of the U.S.-Russia Innovation Corridor (USRIC) ... biotechnology startups to participate in the program since ... the University of Maryland (UMD) signed a Memorandum ... universities in the biomedical industry in 2013. , ...
(Date:4/23/2015)... Calif. , April 23, 2015  Cryoport, ... advanced cryogenic logistics solutions for the life sciences ... established its cryogenic logistics solutions within the animal ... with Yancey Farms to support the shipment of ... German Sport horse Stallions. Animal ...
(Date:4/23/2015)... Proove Biosciences , a commercial ... to announce the launch of Proove Non-Opioid ... 62 genetic variations to predict good versus poor ... non-opioid pain medications, namely ibuprofen (brand name Advil, ... Xanax) and acetaminophen (brand name Tylenol). , ...
(Date:4/23/2015)... 23, 2015 ReliantHeart's HeartAssist5® Ventricular Assist ... flow through the device) in real time, and this ... world’s leading provider of Machine to Machine (M2M) communications ... Flow via VADLink.com can be monitored by ... drop in flow below a minimum threshold, the patient ...
Breaking Biology Technology:U.S.-Russian Innovation Corridor Program Connects Visiting Russian Startups to U.S. Experts 2U.S.-Russian Innovation Corridor Program Connects Visiting Russian Startups to U.S. Experts 3U.S.-Russian Innovation Corridor Program Connects Visiting Russian Startups to U.S. Experts 4Cryoport to Support the Equine Animal Husbandry Market Through Agreement with Yancey Farms 2Cryoport to Support the Equine Animal Husbandry Market Through Agreement with Yancey Farms 3Proove Biosciences Launches World’s First Test to Predict Responders to Commonly Prescribed Non-Opioid Pain Medications 2Proove Biosciences Launches World’s First Test to Predict Responders to Commonly Prescribed Non-Opioid Pain Medications 3
... Palatin Technologies,Inc. (Amex: PTN ) will announce ... 9, 2007 before the open of the U.S. financial,markets. ... live audio webcast,hosted by Palatin,s executive management team on ... of operations in greater detail and provide,an update on ...
... 5 Exelixis,Inc. (Nasdaq: EXEL ) today ... 30, 2007. Revenues for the quarter ended ... million for the comparable period in 2006. The ... due to revenue recognition,associated with new collaboration agreements ...
... patients in over 30 states on ... options since 1997, DEERFIELD, Ill., Nov. 5 ... Kidney Patient,Educator (KPE) program, an ambitious project that has ... treatment options to,nearly 80,000 kidney disease patients in over ...
Cached Biology Technology:Palatin Technologies to Report Fiscal Year 2008 First Quarter Results; Teleconference and Webcast on November 9, 2007 2Exelixis Announces Third Quarter 2007 Financial Results and Business Update 2Exelixis Announces Third Quarter 2007 Financial Results and Business Update 3Exelixis Announces Third Quarter 2007 Financial Results and Business Update 4Exelixis Announces Third Quarter 2007 Financial Results and Business Update 5Exelixis Announces Third Quarter 2007 Financial Results and Business Update 6Exelixis Announces Third Quarter 2007 Financial Results and Business Update 7Exelixis Announces Third Quarter 2007 Financial Results and Business Update 8Baxter Kidney Patient Education Program Celebrates Tenth Anniversary 2Baxter Kidney Patient Education Program Celebrates Tenth Anniversary 3Baxter Kidney Patient Education Program Celebrates Tenth Anniversary 4
(Date:4/1/2015)... , April 1, 2015   Medisafe ™, the ... users across iOS and Android ... integrating biofeedback into the platform to allow patients to ... For the first time, patients will be able to ... dose impacts important biometrics, such as glucose levels and ...
(Date:3/31/2015)... --  Guidepoint , a leading global research services firm, ... TRACKER, part of the Guidepoint TRACKER suite ... device and therapeutics markets. The Post-Surgical Pain Management TRACKER ... share, and adoption rates across three major modalities used ... The Post-Surgical Pain Management TRACKER will provide length ...
(Date:3/26/2015)... TORONTO , March 26, 2015 ... premier family, social, recreation and athletic club, today announced ... designed to allow freedom of movement for members and ... "Following a comprehensive process, we selected FST,s IMID ... sophistication and convenience for our members and staff, in ...
Breaking Biology News(10 mins):Medisafe Announces Launch of Biometric Integration, Enabling Patients to Stay Adherent to Their Meds and See Tangible Health Results 2Guidepoint Launches Post-Surgical Pain Management TRACKER 2Guidepoint Launches Post-Surgical Pain Management TRACKER 3Granite Club First Private Club in North America to Implement FST Biometrics Secure Access Solution, as Part of Multi-million Dollar Expansion 2
... hard to come by, a University of Central Florida and ... National Institutes of Health highly competitive grants for hemophilia research. ... is aimed at determining whether a green technique pioneered at ... The second grant worth $2 million for four years is ...
... The Newark College of Engineering (NCE) at NJIT has been ... one of 20 universities and colleges across the nation in ... programs. This selection is the culmination of a ... Associate Dean Lisa Axe. Currently, some 43 percent ...
... out of the lab for a test drive with ... new way to develop possible future scenarios. James ... Center for Systems Integration and Sustainability (CSIS) and now ... London, has paired the relatively new use of agent-based ...
Cached Biology News:Hemophilia research gets NIH boost to a tune of $5.5 million 2Hemophilia research gets NIH boost to a tune of $5.5 million 3NSF funds NJIT's participation in program to retain engineering students 2Engaging land-use stakeholders is model behavior 2
... a modern flow cytometer, a scientist can ... labeled with fluorescent probes. However, the emission ... cell labeling are broader than the light ... filters. Thus, the fluorescence signal of a ...
... Shaker ,Product Information ... a high quality, general purpose shaker with economical ... other models in the line, the Orbit LS ... panel. Electronic controls are utilized for setting speed ...
... , ,Product Information , ,The ... GX-1000 is extremely efficient, yet gentle, making ... well as staining gels, hybridization and other ... a broad range. A nonslip rubber mat ...
Large-capacity refrigerated incubator chamber holds up to 4 independently-operated shakers. Programmable heat, refrigeration, humidifcation are standard. Photosynthetic operation, optional....
Biology Products: