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Biochemistry JoVE (Open Access) Citable · DOI

Silk Film Culture System for in vitro Analysis and Biomaterial Design

DOI: 10.3791/3646-v
What you'll learn
  • ✓ Fabricate customizable silk films using silicone rubber molds
  • ✓ Prepare biocompatible silk solution from raw silk fibroin
  • ✓ Establish in vitro cell culture system on silk film substrates
  • ✓ Evaluate cell growth and viability on silk biomaterials
Protocol

Silk films are a novel class of biomaterials readily customizable for an array of biomedical applications. The presented silk film culture system is highly adaptable to a variety of in vitro analyses. This system represents a biomaterial design platform offering in vitro optimization before direct translation to in vivo models.

Difficulty
intermediate
Total time
~3–4 days (including silk processing, film preparation, and multi-day culture analysis)
Model organism
HCLE (human corneal limbal epithelial cells)
Biosafety
BSL-1

Steps

1
Fabricate silicone rubber molds for silk films

Create custom silicone rubber molds that serve as templates for silk film casting. These reusable molds enable consistent, reproducible silk film geometry for standardized in vitro culture.

▶ 01:30
2
Produce silk fibroin solution from raw silk

Extract and process silk fibroin protein from cocoons to generate a biocompatible solution. This solution serves as the base material for casting silk films with controlled properties.

▶ 03:11
3
Prepare silk films and establish culture system

Cast silk solution into molds and allow films to dry. Set up the prepared silk films as substrate for cell culture, including sterilization and mounting in culture wells.

▶ 06:32
4
Seed and culture HCLE cells on silk films

Plate human corneal limbal epithelial cells onto prepared silk film substrates and monitor cell adhesion, proliferation, and morphology over culture duration to assess biocompatibility.

▶ 09:03
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