Extensive characterization of feline intra-abdominal adipose-derived mesenchymal stem cells

Abstract

Mesenchymal stem cells (MSCs) isolated from various tissues have been well characterized for therapeutic application to clinical diseases. However, in contrast to MSCs from other animal species, the characteristics of feline MSCs have not been fully documented. In this study, we conducted extensive characterization of feline adipose tissue-derived MSCs (fAD-MSCs). Study fAD-MSCs were individually isolated from the intra-abdominal adipose tissues of six felines. The expression levels of cell surface markers and pluripotent markers were evaluated. Next, proliferation capacity was analyzed by performing cumulative population doubling level (CPDL) and doubling time (DT) calculation assays. Differentiation potentials of fAD-MSCs into mesodermal cell lineages were analyzed by examining specific staining and molecular markers. All fAD-MSCs positively expressed cell surface markers such as CD29, CD44, CD90, CD105, CD166, and MHC-I, while CD14, CD34, CD45, and CD73 were negatively expressed. The CPDL of the fAD-MSCs was maintained until passage 5 to 6 (P5 to P6), whereas DT increased after P3 to P4. Also, stem cell-specific pluripotent markers (Oct3/4, Nanog, and SSEA-4) were detected. Importantly, all fAD-MSCs demonstrated mesodermal differentiation capacity. These results suggest that fully characterized fAD-MSCs could be beneficial when considering the use of these cells in feline disease research.

Viability, Yield and Expansion Capability of Feline MSCs Obtained from Subcutaneous and Reproductive Organ Adipose Depots

“…These samples also had the slowest P2 doubling time. The range of doubling times reported here are consistent with some previous publications of feline abdominal adipose tissue [28][29][30] but slower than those reported by others [20,31].…”

“…However, the presence of Sox2 in feline adipose-derived MSCs is controversial. Some have reported the inability to detect Sox2 or the resulting protein in feline abdominal or subcutaneous adipose MSCs [28,29]. In contrast, our results are consistent with those of Lee et al, showing detection of Sox2 in the abdominal adipose tissue [31].…”

“…In felines, there have been few studies to compare sources of MSCs and they have focused on comparing bone marrow-derived MSC to adipose MSCs [19]. Further, when working with abdominal adipose tissue, the exact donor site often is not clarified in the reports [28,29].…”

A solution to prevent secondary flow in adherent cell cultures

“…Challenging the paradigm of the constitutive representation of MSC-markers, research shows that CD105 expression can be controlled by oxygen availability in the cellular micromilieu (Sánches-Elsner et al, 2002; Mark et al, 2013), while CD90 representation on the cell surface can depend on the extent of cell–cell connections (Kim et al, 2017). In both cases, these markers have a negative correlation, respectively, with their above-mentioned parameters.…”

Isolation and preservation of multipotent mesenchymal stem cells from bone marrow of Arabian leopard (<i>Panthera pradus nimr</i>)

“…Currently, adipose-derived MSCs (aMSCs) are most commonly used in clinical applications due to ease of attainment and their superior proliferative ability (Webb et al, 2012). Adipose tissue is usually collected from ventral abdomen just caudal to the umbilicus by 8 mm punch biopsy or peritoneal fat is collected during spay procedure (Kim et al, 2017). Authors previous experience showed that even a small surgical wound might bring a risk of severe complications.…”