Version 1
: Received: 27 February 2020 / Approved: 28 February 2020 / Online: 28 February 2020 (12:24:54 CET)
Version 2
: Received: 12 March 2020 / Approved: 12 March 2020 / Online: 12 March 2020 (13:24:46 CET)
How to cite:
Kadam, S.; Vandana, M.; Kaushik, K. To Serum or Not to Serum: Reduced-serum Based Approaches for Contact-based Co-culture of Fibroblasts and Keratinocytes for Wound Bed Studies. Preprints2020, 2020020430. https://doi.org/10.20944/preprints202002.0430.v2
Kadam, S.; Vandana, M.; Kaushik, K. To Serum or Not to Serum: Reduced-serum Based Approaches for Contact-based Co-culture of Fibroblasts and Keratinocytes for Wound Bed Studies. Preprints 2020, 2020020430. https://doi.org/10.20944/preprints202002.0430.v2
Kadam, S.; Vandana, M.; Kaushik, K. To Serum or Not to Serum: Reduced-serum Based Approaches for Contact-based Co-culture of Fibroblasts and Keratinocytes for Wound Bed Studies. Preprints2020, 2020020430. https://doi.org/10.20944/preprints202002.0430.v2
APA Style
Kadam, S., Vandana, M., & Kaushik, K. (2020). To Serum or Not to Serum: Reduced-serum Based Approaches for Contact-based Co-culture of Fibroblasts and Keratinocytes for Wound Bed Studies. Preprints. https://doi.org/10.20944/preprints202002.0430.v2
Chicago/Turabian Style
Kadam, S., Madhusoodhanan Vandana and Karishma Kaushik. 2020 "To Serum or Not to Serum: Reduced-serum Based Approaches for Contact-based Co-culture of Fibroblasts and Keratinocytes for Wound Bed Studies" Preprints. https://doi.org/10.20944/preprints202002.0430.v2
Abstract
Contact-based co-culture of fibroblasts and keratinocytes is important to study the structure and functions of the wound bed. Co-culture of these two cell types in direct contact with each other has been challenging, requiring high serum concentrations (up to 10%), feeder systems and a range of supplemental factors. These approaches are not only technically demanding, but also present scientific, cost and ethical limitations associated with high-serum concentrations. We have developed two reduced-serum approaches (1-2%) to support contact-based co-culture of human dermal fibroblasts (HDFa) and human epidermal keratinocytes (HaCaT). The two approaches include (1) Specialized cell culture media for each cell type mixed in a 1:1 ratio (KGM+FGM), and (2) Minimal media supplemented with cell-specific growth factors (MEM+GF). Co-culture could be successfully achieved by co-seeding (two cell types were introduced simultaneously), or in a layered fashion (keratinocytes seeded on top of confluent fibroblasts). With wound scratch assays, the co-cultured platforms could demonstrate cell proliferation, migration and wound closure. The reduced-serum conditions developed are simple, easy to formulate and adopt, and based on commonly-available media components. These contact-based co-culture approaches can be leveraged for wound and skin studies, and tissue bioengineering applications, potentially reducing concerns with high-serum formulations.
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Commenter: Karishma Kaushik
Commenter's Conflict of Interests: Author