Nippi Research Institute of Biomatrix 1 article published in JoVE Bioengineering Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates Hitomi Fujisaki1, Sugiko Futaki2, Kazunori Mizuno1, Shunji Hattori1 1Nippi Research Institute of Biomatrix, 2Department of Anatomy and Cell Biology, Osaka Medical College Here, we present a protocol for the preparation of two different forms of culture substrates utilizing type I collagen. Depending on how collagen is handled, collagen molecules either maintain two-dimensional, non-fibrous form or reassemble into three-dimensional, fibril form. Cell proliferation on type I collagen is drastically affected by fibril formation.