Browsing by Person "Nellinger, Svenja"

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Animal‐free setup of a 3D mature adipocyte‐macrophage co‐culture to induce inflammation in vitro
(2025) Nowakowski, Sophia; Nellinger, Svenja; Albrecht, Franziska Brigitte; Kluger, Petra Juliane
Adipose tissue inflammation plays a central role in the pathogenesis of metabolic disorders. It is closely associated with immune cell infiltration, particularly macrophages, and the release of pro‐inflammatory cytokines. Reliable in vitro test systems that mimic the inflamed environment while being free of animal‐derived components are essential to explore new treatments for obesity‐related diseases. This study aims to develop a straightforward, animal‐free adipocyte‐macrophage co‐culture for investigating adipose tissue inflammation. Therefore, the human monocytic cell lines Mono Mac (MM6) and THP‐1 are co‐cultured with human primary mature adipocytes (ACs) encapsulated in gellan gum (GG) within a defined environment. Both monocytic cell lines are effectively activated by phorbol 12‐myristate 13‐acetate (PMA) and lipopolysaccharide (LPS) in the defined medium, exhibiting distinct cytokine profiles. A comparison between collagen and GG demonstrates that GG is a suitable animal‐free matrix material for ACs. PMA+LPS successfully activates the 3D adipocyte‐macrophage co‐culture to an inflammatory state for 72 h in the developed defined medium. Viability and intracellular lipid content remain high, and the functionality of ACs (perilipin A) in untreated models remains intact. This inflamed adipocyte‐macrophage co‐culture is easy to assemble and set up in a defined environment, making it a potential test system for anti‐inflammatory treatment strategies.
Dairy byproducts as sustainable alternatives to FCS in 2D and 3D skeletal muscle cell cultures
(2025) Baldeweg, Tobias Horst; Hubel, Philipp; Günther, Johannes; Ostertag, Fabian; Nellinger, Svenja; Heine, Simon; Kluger, Petra Juliane
Skeletal muscle tissue engineering is a rapidly developing field with applications in disease modelling, tissue replacement, biorobotics, and cultivated meat. The need for more sustainable and ethical biotechnologies has grown due to concerns about resource scarcity, climate change, and animal welfare. One major challenge is replacing fetal calf serum (FCS), a widely used but ethically and environmentally highly problematic media supplement. A promising alternative is the utilization of natural byproducts such as whey and colostrum from the dairy industry, which provide essential nutrients and growth factors. In this study, wheys produced by microfiltration of raw milk and colostrum were investigated as FCS replacements for culturing C2C12 skeletal muscle cells. Composition analysis confirmed a variety of pro-proliferative compounds in both substances. Cell culture experiments led to the development of an optimized medium formulation based on colostrum whey. Colostrum whey medium (CM) supported cell proliferation and maintained the myogenic differentiation potential for over 30 days. Additionally, a CM-based freezing solution enabled effective cryopreservation throughout culture. In 3D static suspension culture, CM sustained viable spheroids for over 14 days. Spheroids showed significantly higher proliferation compared to those in serum-containing medium, making CM suitable for 3D modelling and scale-up of biomass production. These findings highlight CM as a sustainable, cost-effective, and ethical alternative for skeletal muscle tissue engineering, particularly in cultivated meat production.
Gellan gum is a suitable biomaterial for manual and bioprinted setup of long-term stable, functional 3D-adipose tissue models
(2022) Albrecht, Franziska B.; Dolderer, Vera; Nellinger, Svenja; Schmidt, Freia F.; Kluger, Petra J.
Due to its wide-ranging endocrine functions, adipose tissue influences the whole body’s metabolism. Engineering long-term stable and functional human adipose tissue is still challenging due to the limited availability of suitable biomaterials and adequate cell maturation. We used gellan gum (GG) to create manual and bioprinted adipose tissue models because of its similarities to the native extracellular matrix and its easily tunable properties. Gellan gum itself was neither toxic nor monocyte activating. The resulting hydrogels exhibited suitable viscoelastic properties for soft tissues and were stable for 98 days in vitro. Encapsulated human primary adipose-derived stem cells (ASCs) were adipogenically differentiated for 14 days and matured for an additional 84 days. Live-dead staining showed that encapsulated cells stayed viable until day 98, while intracellular lipid staining showed an increase over time and a differentiation rate of 76% between days 28 and 56. After 4 weeks of culture, adipocytes had a univacuolar morphology, expressed perilipin A, and secreted up to 73% more leptin. After bioprinting establishment, we demonstrated that the cells in printed hydrogels had high cell viability and exhibited an adipogenic phenotype and function. In summary, GG-based adipose tissue models show long-term stability and allow ASCs maturation into functional, univacuolar adipocytes.