BioNOC II^® Cell Culture Macrocarriers

Features

• Productivity: Up to 2-3X10⁹ cells/0.1 L of BioNOC II^® II (for Vero cells)
• Large growth surface area: Up to 2,730 cm² /g of BioNOC II^® II
• High surface area to volume ratio: 160 cm²/ml
• 100% Polyethylene Terephthalate (PET) material
• 0.3 mm thickness for easy exposure to culture medium oxygen transfer
• Non-pyrogenic, non-toxic, animal component-free
• Special surface treatment to enhance biocompatibility
• Special 3D geometric design to enhance mixing and support load-bearing capacity in packed-bed bioreactors
• Compressible for higher cell density
• Hydrophilicity stable for up to four years
• Sterilizable by autoclave (if not pre-packed), gamma irradiation, and EO sterilization
• Extremely low lint waste, convenient for adherent cell growth for protein or viral vaccine production

Highlights

1. 1. Mimics the cell’s in vivo environment

   Various anchorage-dependent cell lines such as Chinese Hamster Ovary (CHO), Kidney Epithelial Cell (VERO), Madin-Darby Canine Kidney Cell (MDCK), and Insect Cell (SF9) were used for vaccine, protein, and monoclonal antibody productions. The use of roller bottles and cell factories were prominent for the large-scale cell culture of these adherent cells; however, the development was costly, expensive, and always at risk of contamination.

   Microcarrier-based cell cultures have since made use of stirred tank bioreactors (STR) and fixed-bed bioreactors with macrocarriers. Compared with STR, the fixed-bed bioreactor has the capacity to protect cells from stressful conditions and perfusion culture model is easily achievable. Singapore-based macroporous carrier manufacturer Esco VacciXcell, supplies BioNOC II^® for use with different cell types. The use of macroporous carriers is a promising technique that has advantages in enhancing production capacity, improving the culture’s robustness, and facilitates process scale-up allowing the culture of various animal cells in high yield.

   Unlike other types of microcarrier for cell culture, a macroporous carrier such as BioNOC II^® creates a multilayer culture (3D cell growth) that allows cell to cell and cell to extracellular matrix interactions, mimicking the cell's in vivo environment. This is highly essential for multiple cellular processes, including differentiation and proliferation. Materials used in the creation of BioNOC II^® are FDA-approved, cGMP compliant, and this macroporous carrier manufacturing passed regulation guidelines USP Class VI, USP <87>, <83>, ISO 10993-5.

   Vero Cells:

   Live cells (fluorescent) staining of Vero cells on BioNOC II^® II under 4x magnification. Calcein green staining of cytoplasm and Hoechst 33342 (blue) staining of the nucleus.

   Mesenchymal Stromal Cells:

   Black arrow points show ECM CL-MSCs cultured 4 days in BioNOC II^® II - staining of collagen fibers with picro sirius red

2. 2. Geometric design and surface treatment

   This macroporous carrier also has a distinct geometrical design that is specially folded to avoid overlapping of cells. This allows sufficient nutrient and oxygen transfer.

   Adherent cells attached to BioNOCTM II carriers are alternately exposed to aeration and nutrition phases by pumping media in and out of the packed bed.

   BioNOC II^® II also exhibits great surface treatment. This macrocarrier can have a stable hydrophilicity for up to 4 years. It is non-cytotoxic that passed both serum and serum-free medium tests. Some cells are loosely adherent in nature, which is why they require with coating factors prior to cultivation to support higher attachment efficiency. However, not all microcarriers can support this. This limitation is being addressed with the use of BioNOC II^® II. Due to its enhanced hydrophilicity, the addition of coating factors is easily accommodated.

3. 3. Large Surface Area for Adherent Cell Growth

   BioNOC II^® II presents straight and uniform fibers which are highly favored for cell distribution and growth. It also possesses higher density that prevents it from floating during the cell culture and remains stable all throughout the culture run.

   Vero cells have been cultured in BioNOC II^® II and are grown to high densities from 2-3x109 using batch or perfusion mode of culture using Tide Motion bioreactors such as CelCradleTM and TideXcellTM. The surface area and the cell numbers achieved for Vero cell culture are shown to be very promising. These results prove that the BioNOC II^® II macrocarrier allows for a robust linear scalability, and thus saves time, labor, and resources for process development.

   Day 6:

   Day 14:

   Figure 1. Live cell fluorescent staining of Vero cells on BioNOCTM II under 4x magnification. Calcein green staining of cytoplasm, Hoechst 33342 (blue) staining of the nucleus and Propidium Iodide stain for dead cells was done.

   The fold expansion of Vero cells from attachment on BioNOCTM II in a 1L packed bed to harvest using an automated TideXcell Harvesting System (TXLHS) was 19-fold and the efficiency of harvest was 93.6%.

   Ten (10) pieces of BioNOCTM II macrocarriers is equivalent to one (1) T-25 flask.

   

   Table 1. Cell growth in SFM of different carriers
   Model	Cell Density
   Microcarriers	2.6 x 106 cells/ml
   Macrocarriers	5.8 x 106 cells/ml

   Table 2. Cell growth in serum-containing media of different carriers
   Model	Cell Density
   Microcarriers	3.3 x 106 cells/ml
   Macrocarriers	6.5 x 106 cells/ml

   Comparisons
   	BioNOC II® II	Commercial Disks
   Material	100% PET	PET / PP
   Dimension	55 mm x 10 mm strip	Disk diameter 6 mm
   Specific Surface area	2,400	1,200
   Packed volume (ml/g)	15	10
   S/V	160	120
   Autoclavable	OK	OK
   Gamma irradiation	OK
   Endotoxin tested (EU/ml)	< 0.25	< 0.5
   Bioburden (CFU/gram)	< 1	< 0.36
   Cytotoxicity tested	Serum and serum-free culture test	Yes
   Regulation guideline	USP <87> <88>, ISO10993-5	USP <87> <88>, ISO10993-5