BRIEF RESEARCH REPORT article
Front. Immunol.
Sec. Molecular Innate Immunity
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1589553
Optimizing Monocyte-Derived Immune Cell Cultures: Comparing Xeno-Free and Xenogeneic Conditions
Provisionally accepted
- 1Department of Clinical Dentistry, Faculty of Medicine, University of Bergen, Bergen, Hordaland, Norway
- 2Haukeland University Hospital, Bergen, Hordaland, Norway
- 3Department of Clinical Medicine 2, Faculty of Medicine, University of Bergen, Bergen, Hordaland, Norway
- 4Department of Clinical Medicine 1, Faculty of Medicine, University of Bergen, Bergen, Hordaland, Norway
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Culture conditions significantly affect the phenotype of immune cells. This study compared, for the first time, the impact of xeno-free human AB serum and xenogeneic fetal bovine serum (FBS) culture conditions on the surface marker expression of monocyte-derived macrophages (Mo-Mø) and dendritic cells (Mo-DC) using spectral flow cytometry. Monocytes were differentiated into macrophages or Mo-DC over 6 days. M0 macrophages were polarized toward M1-like or M2-like macrophages, and Mo-DC were activated using LPS. Differentiation was successful in both conditions. Despite cells exhibiting autofluorescence, distinct phenotypes based on selected differentiation markers were observed. CD1a expression was lacking in AB cultures, while expression of CD16, CD163 and the co-stimulatory molecules CD80 were significantly upregulated and CD86 downregulated by xenogeneic FBS conditions. These novel findings highlight the need for careful selection of serum type and phenotyping markers to minimize unexpected results and account for potential serum-induced interference with marker expressions in monocyte-derived immune cells.
Keywords: dendritic cell, Human AB serum, Fetal bovine serum, macrophage, Spectral flow cytometry
Received: 07 Mar 2025; Accepted: 25 Sep 2025.
Copyright: © 2025 Marek, Wolff, Dongre and Suliman. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Salwa Suliman, salwa.suliman@uib.no
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