Overview
Colossal Biosciences, the biotech company known for its ambitious de-extinction projects, has developed a groundbreaking artificial eggshell that allows chicken embryos to develop outside a natural egg. This technology, which uses a 3D-printed plastic lattice coated with a specialized silicone membrane, mimics the oxygen permeability of a real eggshell and provides a transparent window for observation. The innovation is a key step toward resurrecting extinct avian species like the dodo and the giant moa, as well as aiding conservation of at-risk birds. This guide explains the science behind the artificial eggshell, the step-by-step process of its use, and common pitfalls to avoid.

Prerequisites
Knowledge Requirements
- Basic understanding of avian embryology (e.g., development stages from fertilization to hatching)
- Familiarity with 3D printing materials (e.g., PLA, silicone polymers) and membrane science
- Knowledge of sterile laboratory techniques to prevent contamination
Materials Needed
- 3D printer capable of producing the lattice structure (e.g., FDM or SLA printer)
- Silicone-based membrane material (e.g., PDMS - polydimethylsiloxane) with appropriate oxygen permeability
- Freshly laid chicken eggs (within 24 hours of laying)
- Sterile transfer tools (e.g., pipettes, forceps, sterile Petri dishes)
- Incubator capable of maintaining 37.5°C (99.5°F) and 50–60% humidity
- Transparent observation window (can be pre-integrated into the 3D-printed cup)
Step-by-Step Instructions
Step 1: Fabricate the Artificial Eggshell
Design an oval-shaped lattice structure using CAD software (e.g., Fusion 360). The lattice should have a porosity that allows gas exchange but retains the egg's contents. Print the shell using a biocompatible plastic such as PLA or PETG. Ensure the interior surface is smooth for membrane application.
Step 2: Apply the Silicone Membrane
Mix the silicone-based membrane solution according to manufacturer instructions. Coat the interior of the printed lattice evenly, covering all holes. Cure the membrane under UV light or heat until it forms a thin, flexible layer. The membrane must be gas-permeable (similar to an eggshell's pores) but impermeable to liquids.
Step 3: Prepare the Egg Contents
Use a freshly laid chicken egg. Sterilize the egg surface with 70% ethanol. Gently crack the egg equatorially and carefully pour the contents (yolk, albumen, and embryo) into a sterile container. Avoid breaking the yolk membrane. The embryo should be visible as a small white spot on the yolk.
Step 4: Transfer to Artificial Shell
Using a wide-mouth funnel or sterile pipette, pour the egg contents into the prepared artificial shell. Fill only to the level that mimics natural egg volume (approximately 50 mL for a chicken egg). Seal the top with a clear, gas-permeable cover (e.g., a thin silicone sheet) that includes a small window for observation.
Step 5: Incubate and Monitor
Place the artificial egg in an incubator at 37.5°C with 55% relative humidity. Rotate the egg daily (as natural mother hens do) to prevent the embryo from sticking to the membrane. Observe through the window for signs of development: blood vessels appearing at day 3, movement at day 7, and pipping at day 20–21 for chickens.

Step 6: Handle Hatching
When the chick begins to pip (break the membrane), do not assist unless necessary. Ensure humidity is high (70%) to prevent drying of the chick. After hatching, carefully open the artificial shell and transfer the chick to a brooder with appropriate warmth (35°C for the first week).
Common Mistakes
Membrane Rupture
If the silicone membrane is too thin or applied unevenly, it may tear during transfer. Always test membrane integrity with sterile water before introducing the embryo.
Contamination
Bacterial or fungal contamination can kill the embryo. Use sterile technique throughout: autoclave tools, work under a laminar flow hood, and use antibiotics in the media only if absolutely necessary.
Incorrect Incubation Conditions
Temperature fluctuations above 38°C or below 37°C can cause developmental abnormalities. Use a reliable incubator with a digital thermostat and backup power. Humidity too low will dehydrate the embryo; too high can drown it.
Egg Age
Eggs older than 48 hours at transfer have reduced viability. Use only freshly laid eggs and store them at 15°C if needed, but transfer as soon as possible.
Observation Window Fogging
Condensation can obscure the window. Apply an anti-fog coating or maintain a slight temperature gradient to avoid moisture buildup.
Summary
Colossal Biosciences' artificial eggshell represents a significant advancement in avian reproductive technology, enabling controlled observation and manipulation of embryonic development. By following the steps outlined—fabricating the shell, applying the membrane, transferring the contents, and incubating carefully—researchers can successfully grow chicks outside a natural egg. This technique holds promise for de-extinction projects like the dodo and moa, as well as for conservation of endangered bird species. However, challenges such as membrane integrity and sterility must be addressed to achieve high success rates. As the technology matures, it may become a standard tool in avian biology and conservation.