A Co-Culture System for Studying Cellular Interactions in Vascular Disease

Abstract

Cardiovascular diseases (CVDs) are leading causes of morbidity and mortality globally, characterized by complications such as heart failure, atherosclerosis, and coronary artery disease. The vascular endothelium, forming the inner lining of blood vessels, plays a pivotal role in maintaining vascular homeostasis. The dysfunction of endothelial cells contributes significantly to the progression of CVDs, particularly through impaired cellular communication and paracrine signaling with other cell types, such as smooth muscle cells and macrophages. In recent years, co-culture systems have emerged as advanced in vitro models for investigating these interactions and mimicking the pathological environment of CVDs. This review provides an in-depth analysis of co-culture models that explore endothelial cell dysfunction and the role of cellular interactions in the development of vascular diseases. It summarizes recent advancements in multicellular co-culture models, their physiological and therapeutic relevance, and the insights they provide into the molecular mechanisms underlying CVDs. Additionally, we evaluate the advantages and limitations of these models, offering perspectives on how they can be utilized for the development of novel therapeutic strategies and drug testing in cardiovascular research.

Keywords: cardiovascular diseases; cellular interactions; co-culture models; endothelial dysfunction; vascular pathology.

Figure 1

Schematic representation of the multi-cellular components of the myocardial environment. Atherosclerotic lesions consist of three important components: a fibrous part composed of extracellular lipids and connective tissue matrix, a cellular part composed of immune cells (monocytes, macrophages), and smooth muscle cells and foam cells, which are formed by the accumulation of lipids in macrophages. Cardiomyocytes associate with the neighboring cells such as endothelial cells and fibroblasts and aid in their cellular function. Immune cells adhere to the vascular tissue upon disease condition, and the macrophages engulf lipoproteins to form foam cells, which accelerates atherosclerosis.

Figure 2

Illustration of the different types of 2D cell culture. (A) Formation of monolayer by a single type of cell in the Petri dish. (B) Direct contact co-culture model: (i) Two different types of cells are mixed at a standard ratio and inoculated on the same culture dish, and (ii) a trans-well-based model where each cell type is cultured to the opposite sides of the permeable membrane in the trans-well chamber. This type of culture ensures physical contact and juxtacrine and paracrine signaling, and these cells tend to maintain their function and structure. (C) Indirect co-culture model with trans-well insert ensure the cell–cell interactions mediated by secreted factors are released in the culture media. (D) Conditioned media-based co-culture system, where the cellular secretions from one cell type are transferred to the other cell type.

Figure 3

Advanced co-culture systems used in vascular disease. (A) Scaffold (hydrogel)-based co-culture models mimic the extracellular matrix and are used to study cell migration and proliferation. (B) Microfluidic co-culture system encompasses a dynamic culture environment (shear stress of the blood flow) to the co-cultured cells to study the cell physiology and drug testing. (C) Heterotypic 3D spheroids consist of multiple cell types that can mimic in vivo physiology.