Abstract
DEVELOPMENT OF A CO-CULTURE SYSTEM FOR EXAMINATION OF ADIPOSE MACROPHAGE CROSS-TALK
Cytotherapy (Oxford, England), Vol.27(5 Supplement), pp.S106-S107
05/2025
DOI: 10.1016/j.jcyt.2025.03.206
Abstract
Adipose tissue is a potential source of cells for several types of cell therapy including MSCs, adipocytes, and macrophages. However, adipose tissue cellular composition and the function of the cells within adipose is heavily influenced by the metabolic health of the individual. With obesity rates reaching epidemic levels in the United States, it is important to understand how changes in metabolic state impact the function of adipose derived cells. To facilitate this work, we sought to create an in vitro model of adipose that incorporates adipocytes, MSCs, and macrophages. Our aim was to develop a co-culture technique to identify the influence of macrophage-adipocyte crosstalk on mature adipocyte function, adipose-derived MSC potential, and adipocyte influences on macrophage phenotype.
Stromal vascular fraction from surgical adipose tissue was culture expanded and characterized for MSC markers. MSC organoids were formed via agarose micromolds containing 35 microwells to form 10,000 cell organoids. The organoids were differentiated with adipogenic media for 14 days. Human monocytes were isolated from leukocyte reduction cones, differentiated into naïve macrophages, and polarized to either pro-inflammatory (M1-like) or anti-inflammatory (M2-like) states. To study mature adipocyte-macrophage cross-talk, micromolds were transferred to macrophage wells, cultured within agarose punchouts to control for macrophage seeding density and cell ratios. Media was collected for secretome analysis, and spheroids were stained and imaged through confocal microscopy for lipid content.
The secretome of macrophage/adipocyte co-cultures show distinct profiles between each cell type, and therefore, inflammatory environment. While both macrophage types caused diminished adiponectin production by adipocytes along with induced lipolysis without the presence of catecholamines, the presence of adipocytes lead to a decrease of pro-inflammatory cytokines with M2-like macrophages, yet an increase with M1s.
This work demonstrates the impact of adipose health on the quality of cells for cell therapies. Chronic inflammation impairs adipogenesis and impacts all the cells within adipose. Future studies need to be performed to identify the influence of inflammatory environment on MSC health, along with repeating further studies with additional donors.
Details
- Title: Subtitle
- DEVELOPMENT OF A CO-CULTURE SYSTEM FOR EXAMINATION OF ADIPOSE MACROPHAGE CROSS-TALK
- Creators
- J. Liszewski - University of IowaR. Behan-Bush - University of IowaY. Wan - Biomedical Engineering, The University of Iowa Roy J Carver Department of Biomedical Engineering, Iowa City, IA, United StatesA. Klingelhutz - University of Iowa, Microbiology and ImmunologyE. Sander - University of IowaJ.A. Ankrum - University of Iowa
- Resource Type
- Abstract
- Publication Details
- Cytotherapy (Oxford, England), Vol.27(5 Supplement), pp.S106-S107
- DOI
- 10.1016/j.jcyt.2025.03.206
- ISSN
- 1465-3249
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 05/2025
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Microbiology and Immunology; Iowa Technology Institute; Orthopedics and Rehabilitation; Radiation Oncology; Craniofacial Anomalies Research Center; Fraternal Order of Eagles Diabetes Research Center; Chemical and Biochemical Engineering
- Record Identifier
- 9984820570802771
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