Biologically relevant laminin as chemically defined and fully human platform for human epidermal keratinocyte culture

Monica Suryana Tjin, Alvin Wen Choong Chua (), Aida Moreno-Moral, Li Yen Chong, Po Yin Tang, Nathan Peter Harmston, Zuhua Cai, Enrico Petretto, Bien Keem Tan and Karl Tryggvason ()
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Monica Suryana Tjin: Duke-NUS Medical School
Alvin Wen Choong Chua: Singapore General Hospital
Aida Moreno-Moral: Duke-NUS Medical School
Li Yen Chong: Duke-NUS Medical School
Po Yin Tang: Singapore General Hospital
Nathan Peter Harmston: Duke-NUS Medical School
Zuhua Cai: Duke-NUS Medical School
Enrico Petretto: Duke-NUS Medical School
Bien Keem Tan: Singapore General Hospital
Karl Tryggvason: Duke-NUS Medical School

Nature Communications, 2018, vol. 9, issue 1, 1-10

Abstract: Abstract The current expansion of autologous human keratinocytes to resurface severe wound defects still relies on murine feeder layer and calf serum in the cell culture system. Through our characterization efforts of the human skin basement membrane and murine feeder layer 3T3-J2, we identified two biologically relevant recombinant laminins—LN-511 and LN-421- as potential candidates to replace the murine feeder. Herein, we report a completely xeno-free and defined culture system utilizing these laminins which enables robust expansion of adult human skin keratinocytes. We demonstrate that our laminin system is comparable to the 3T3-J2 co-culture system in terms of basal markers’ profile, colony-forming efficiency and the ability to form normal stratified epidermal structure in both in vitro and in vivo models. These results show that the proposed system may not only provide safer keratinocyte use in the clinics, but also facilitate the broader use of other cultured human epithelial cells in regenerative medicine.

Date: 2018
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DOI: 10.1038/s41467-018-06934-3

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