Preprint Article Version 2 This version is not peer-reviewed

To Serum or Not to Serum: Reduced-serum Based Approaches for Contact-based Co-culture of Fibroblasts and Keratinocytes for Wound Bed Studies

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. Preprints 2020, 2020020430 (doi: 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 (doi: 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.

Subject Areas

Co-culture; Reduced-serum; Wound bed; Fibroblasts; Keratinocytes

Comments (1)

Comment 1
Received: 12 March 2020
Commenter: Karishma Kaushik
Commenter's Conflict of Interests: Author
Comment: Extensive edits to the writing and review of literature.
+ Respond to this comment

We encourage comments and feedback from a broad range of readers. See criteria for comments and our diversity statement.

Leave a public comment
Send a private comment to the author(s)
Views 0
Downloads 0
Comments 1
Metrics 0


×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.