Culturing Cells in Defined 3-D Structures

Media Supplements Matter

There have been several publications referencing use of our growth factors in 3-D Cultures. It is important that potent growth factors are used to ensure proper cell growth and differentiation.

Here's a new publication referencing use of our ISOKine^TM FGF. Our ISOKine growth factors are 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.

Claas Willem Visser, Tom Kamperman, Lisanne P. Karbaat, Detlef Lohse and Marcel Karperien. In-air microfluidics enables rapid fabrication of emulsions, suspensions, and 3D modular (bio)materials. Science Advances 31 Jan 2018: Vol. 4, no. 1, eaao1175 DOI: 10.1126/sciadv.aao1175. In this study, the authors present in-air microfluidics (IAMF), a new chip-free platform technology that enables in-flight (that is, on-the-fly) formation of droplets, fibers, and particles and their one-step deposition into 3D constructs with a modular internal architecture.

Figure: Concept of IAMF and guide to the article. (A) Chip-based microfluidics enables in-line control over droplets and particles, making it a versatile platform technology. A chip design where droplets (blue) are transported by a coflow (pink) is shown. (B) IAMF maintains the in-line control of chip-based microfluidics but relies on jet ejection and coalescence into air. Therefore, a wide range of droplets and particles can be produced at flow rates typically two orders of magnitude higher than with chip-based microfluidics. When combining reactive, solidifying microjets, IAMF also enables on-the-fly production and direct deposition of microparticles into 3D multiscale modular (bio)materials.

Figure: One-step additive manufacturing and injection molding of 3D multiscale modular (bio)materials. (A) Modular free forms with a controlled microarchitecture were manufactured by stacking of shape-stable core-shell particles. (B to D) A hollow cylinder was formed by deposition of the composite jet onto a rotating substrate. By altering the building blocks’ composition, the resulting microarchitecture consisted of (C) a liquid-filled foam or (D) a multimaterial modular solid, where the cross-linker for the core was added to the shell and vice versa. (E) To eject a modular filler, only the droplets’ cores are solidified in the air, whereas the slower solidifying shells enable seamless filling of the mold. (F to H) A modular construct was produced by filling a bone-shaped mold. Inset: Hydrogel construct while still in the mold. The 3D multiscale modular material consisted of MSCs (pink), encapsulated in alginate microspheres (green) that are embedded in dextran-tyramine hydrogel (red). (I) Injection-molded multiscale modular tissue construct with optimized cellular micro- and macroenvironments. The construct consisted of insulin-producing pancreatic β cells (MIN6; beige with blue nuclei) that were encapsulated in alginate microparticles (green). The cell-laden microparticles were encapsulated within a proangiogenic fibrin network that contained human endothelial and stem cells (pink with blue nuclei). The microenvironments supported MIN6 cell proliferation, whereas the macroenvironment supported the formation of an endothelial cellular network within 7 days of in vitro culture. HUVEC, human umbilical cord endothelial cell. Scale bars, 1 cm (B and F), 5 mm (G), and 100 μm (C, D, H, and I).

We live in a 3-D world and 3-D Cell and Tissue Based Assays are a major focus for us. This includes bioinks for 3-D printing.