Mesenchymal Stem Cell Exosomes Derived from Feline Adipose Tissue Enhance the Effects of Anti-Inflammation Compared to Fibroblasts-Derived Exosomes

Abstract

Adipose tissue-derived mesenchymal stem cells (AD-MSCs) release extracellular vesicles such as exosomes, apoptotic bodies, and microparticles. In particular, exosomes are formed inside cells via multivesicular bodies (MVBs), thus their protein, DNA, and RNA content are similar to those of the parent cells. Exosome research is rapidly expanding, with an increase in the number of related publications observed in recent years; therefore, the function and application of MSC-derived exosomes could emerge as cell-free therapeutics. Exosomes have been isolated from feline AD-MSCs and feline fibroblast cell culture media using ultracentrifugation. Feline exosomes have been characterized by FACS, nanoparticle tracking analysis, and transmission electron microscopy imaging. Moreover, cytokine levels were detected by sandwich enzyme-linked immunosorbent assay in exosomes and LPS-induced THP-1 macrophages. The size of the isolated exosomes was that of a typical exosome, i.e., approximately 150 nm, and they expressed tetraspanins CD9 and CD81. The anti-inflammatory factor IL-10 was increased in feline AD-MSC-derived exosomes. However, pro-inflammatory factors such as IL-1β, IL-8, IL-2, RANTES, and IFN-gamma were significantly decreased in feline AD-MSC-derived exosomes. This was the first demonstration that feline AD-MSC-derived exosomes enhance the inflammatory suppressive effects and have potential for the treatment of immune diseases or as an inflammation-inhibition therapy.

Extracellular Vesicles of Mesenchymal Stem Cells Are More Effectively Accessed through Polyethylene Glycol-Based Precipitation than by Ultracentrifugation

“…In the cytokine secretion assay, TNF- α is considered a proinflammatory factor, and IL-10 is an anti-inflammatory factor. THP-1 exposure to LPS resulted in an increase in both, and MSC-EVs reversed this phenomenon and even reduced the level of IL10 [ 44 ]. EVs of equivalent quality derived by UC or PEG function in different modes of secretion inhibition, and the probable causes could be a result of RNA differences in EVs derived by different methods.…”

Extracellular Vesicles Including Exosomes From Animal Mesenchymal Stem/Stromal Cells

“…В нашей работе мы продемонстрировали, что клетки с фенотипом МСК сельскохозяйственных (КРС, овцы, лошадь) и мелких домашних (собака, кошка) животных продуцируют в процессе культивирования ВВ, которые накапливаются в КС, и их можно оттуда извлечь. Ранее экзосомы были получены из МСК лошади (21), собак (22), кошек (23). Для выделения ВВ из КС МСК используется много методов (24)(25)(26), среди которых можно выделить дифференциальное центрифугирование, основанное на разнице в размере и плотности между ВВ и другими веществами, присутствующими в КС.…”

Canine Mesenchymal Stromal Cell Exosomes: State-of-the-Art Characterization, Functional Analysis and Applications in Various Diseases

“…Therefore, products obtained from canine MSCs may also be effective for human treatment. While there have been studies on feline [15] and porcine exosomes [16], the proliferation rate of canine MSCs is higher compared to that of feline and porcine MSCs when supplemented with FBS [17,18]. Hence, over the past decade, not only have the properties of canine MSC-derived exosomes been studied after cell isolation, but also their biological properties.…”