Neuromics' ISOKine bFGF in 3-D Cultures

Works Well in Perfusion Models

Our ISOKine^TM FGF is produced in the endosperm tissue of barley grain (Hordeum vulgare), that exhibits up to 50 times less protease activity than E.coli or mammalian cells. Barley seed is void of any human or animal viral contaminants that could jeopardize your cell culture.

It is a proven solution for all cell cultures and starts at the low price of  65 USD/10 ug.

Here's a reference of its use in 3-D Cultures: Tom Kamperman, Sieger Henke, Claas Willem Visser, Marcel Karperien, Jeroen Leijten. Centering Single Cells in Microgels via Delayed Crosslinking Supports Long-Term 3D Culture by Preventing Cell Escape. DOI: 10.1002/smll.201603711.

Figure: Delayed on-chip crosslinking enables centering of single cells in microgels. a) Fluorescence confocal imaging confirmed that delayed enzymatic crosslinking enabled centering of single MSCs in Dex-TA microgels. b) On average, cell-laden microgels were only 9 µm larger than the encapsulated MSCs, effectively resulting in 3D hydrogel coatings of less than 5 µm. c) A standard microfluidic droplet generator was connected to the H2O2 diffusion-based crosslinking chip. The position of cells (white arrows) in non-crosslinking microgel precursor droplets was analyzed d) immediately after droplet generation (t1), at the start of the crosslinking chip (t2), and e) at the end of the crosslinking chip (t3). f) Cell positions within microgels produced using conventional microfluidic encapsulation systems (i.e., with coupled emulsification and gelation) are indicated with gray (i.e., references) and red (i.e., this work) data points. Cell positions within gel precursor droplets along the modular microfluidic setup are indicated with blue data points. Cell positions within delayed enzymatically crosslinked microgels are indicated with green data points. g) Cell position analyses of various combinations of distinct hydrogel materials (i.e., Dex-TA, Dex-HA-TA, PEGDA), cell types (i.e., MIN6, MSC), and crosslinking methods (i.e., enzyme-based and photo-crosslinking), revealed that delayed crosslinking consistently resulted in significantly increased cell-centering as compared to the conventional encapsulation approach where emulsification and gelation are coupled.

If you are looking for competitively priced, animal free and potent growth factors. check out our ISOkines.

More On Petaka Mini Bioreactors

Getting Culture Conditions Right Every Time

I would like to provide yet more information on the capabilties of our new and innovative Petaka Mini Bioreactors. This posting focuses how oxygen concentrations are tightly controlled in a way that is consistent with the cultured cells natural environment.

The length and cross-section of the respiratory duct is purposely engineered to partially restrict the diffusion of oxygen from the high levels of ambient air to create lower, physiologic levels of dissolved oxygen in the reaction chamber. In accordance to Fick’s Law, as oxygen is consumed inside the culture chamber, decreasing the partial pressure of oxygen in the media, oxygen diffuses in from the outside atmospheric (higher) partial pressure, through the respiratory duct, to the lower partial pressure inside. Diffusion is proportional to the concentration gradient, as regulated by the engineered design of the respiratory duct, and occurs entirely spontaneously and without any manual intervention whatsoever.

Image: Petaka®G3 Ducted Respiratory Chamber (DRC). (1) cell culture chamber;(2) injection port; (3) respiratory duct; (4) 0.2μ filter; (5) water vapor condensers and capillary breakers; (6) unique barcode. The DRC is shown upright in a silicone stand (7).

At the same time, the respiratory duct partially retains carbon dioxide from cellular metabolism to maintain a physiologically normal mild acidosis to balance pH. All gas exchange with the outside environment occurs via a 0.2 micron filtered vent, preserving the internal sterility of the device but allowing exchange of gas diffusion and flow to prevent pressurization issues when filling and emptying the bioreactors.
Therefore, cell culture in DRC’s/Petaka G3 does not require supplemental oxygen-nitrogen balancing, CO2 and humidity sources, eliminating the entire panoply of gas tanks, regulators, sensors, microprocessors and water pans. This creates a double benefit: not only are cells cultured in more normal physiologic conditions, but the mechanics, logistics, risks and costs of cell culture are greatly simplified and reduced.

The Results are Stunning!

Here are result using Petaka® G3 LOT for culturing our UCB Derived hMSCs and Mouse MSCs.

Images: (A) Immuno-fluorescence microscopy of a mouse MSC in differentiation progression. Culture under 20 mmHg of O2 partial pressure. Red fluorescence positive staining of GFAP. hMSC 3 hours after seeding in Petaka G3 with low serum media and without matrix (B) and 96 h later (C). Photos: Jim Musick. Vitro-Biopharma. September, 2012.

Next up using Petaka DRCs for GMP. Stay tuned.