Introducing 2PP 3D printing technology

The collaborative European project INCITE aims to improve current immunotherapies against cancer by manufacturing anti-cancer cells that are better at fighting solid tumours.

To realise this ambitious goal, the INCITE project attempts to create anti-cancer memory T cells that show a high level of proliferation and excellent capabilities of self-renewal and endurance. This means that they can endure the long-lasting battle against a constantly and quickly regenerating tumour.

Generally, anti-cancer T cells originate from “immune niches” that are located within our lymph nodes. However, they are often not mobilized quickly enough, and/or they are produced in insufficient numbers to get rid of the tumour altogether. Therefore, the dual challenge is (I) to grow effective and vital T cells in vitro and (II) to replicate them in large numbers. This is especially needed because the current commercially available T cells show insufficient endurance to overcome solid tumours.

Position of 3D printing technology within the INCITE project

With these objectives in mind, the INCITE project aims to create an artificial immune niche. This immune niche is a carefully constructed environment where T cells can be grown outside the body. Inside the immune niche, the chemical and biological conditions mimic the interior of a lymph node. After all, if we want to grow and replicate T-cells successfully, we must provide them with an environment that is as similar as possible to a human lymph node.

To create this immune niche, we utilize a high-resolution light-based 3D printing technology called two-photon polymerization (2PP). This allows us to rapidly manufacture high-resolution parts. While a regular printer deposits ink in a 2D pattern, a 3D printer enables the fabrication of complex 3D polymer structures.

2PP 3D printing makes manufacturing of extremely small objects possible. This castle rests on the tip of a pencil.

As opposed to conventional manufacturing methods, which work in a subtractive manner, 3D printing is an additive process that enables the rapid fabrication of complex structures.

One of the remarkable aspects of 2PP is its ability to create highly precise and intricate three-dimensional structures that range from the macro-, to the micro-, and nanoscale.

The UpNano 2PP technology extends the printable range from the nanometer (nm) tot the millimeter (mm) range

The 2PP process involves curing a liquid resin in a layer-by-layer fashion, gradually building up a larger and more complex structure. This way of working allows for the formation of structures with intricate geometries, high aspect ratios, and fine feature sizes that are difficult to achieve when using other fabrication techniques.

Video: 2PP allows for polymerizing structures directly inside a sealed microfluidic chip using the bottom-up print mode.

UpNano – one of the partners in the INCITE-project – has developed the fastest 2PP 3D printer on the market, called the NanoOne. By combining high resolution with high throughput, it is possible to manufacture structures at the scale and resolution that is necessary to recreate the lymph node microarchitecture.

For INCITE, UpNano has developed a special resin to print the immune niches. The material is designed to have high transparency and low fluorescence, making it well-suited for confocal imaging. Furthermore, it shows a very good compatibility with living cells, ensuring its suitability for biological applications. The resin itself is liquid, but it turns into a solid polymer when exposed to light by linking its monomer molecules together. The microarchitecture is printed directly inside a sealed microfluidic chip, using the bottom-up printing mode of the NanoOne.

In the next blog, we delve deeper into the specifics of the 3D printing process.

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