Comparison of the Donor Age-Dependent and In Vitro Culture-Dependent Mesenchymal Stem Cell Aging in Rat Model

Abstract

Clinical experiments suggest that mesenchymal stem cells (MSCs) may be useful for tissue repair therapies or treatment of the autoimmune disorders. There is still lack of consensus concerning the age limit of MSC donors, majority of researchers suggest the autologous MSC therapies of patients not exceeding age limit of 55-60 yrs. The purpose of our study was to compare the selected parameters of MSCs from adipose tissue (adipose stem cell, ASC) collected from young and old rats of ages corresponding to patient’s ages 25 yrs. and 80 yrs., respectively. The differences of parameters of ASCs from young and old animals were compared with the differences between ASCs from short-term (3 passage) and long-term (30 passage) in vitro culture. Cell morphology, surface marker expression, growth potential, metabolic activity, β-galactosidase activity, clonogenic potential, angiogenic potential, and differentiation ability of ASCs from young and aged animals and from in vitro cultures at 3rd and 30th passages were compared and analyzed. It may be concluded that ASCs may be applied for autologous transplantations in aged patients. Comparison of ASC aging dynamics depending on host aging or in vitro culture duration suggests that long-term in vitro culture may affect ASCs more than natural aging process of their host. We suggest that ASCs expanded in vitro prior to their clinical use must be carefully screened for the possible aging effects resulting not only from donor age, but from the duration of their in vitro culture.

A Comparative Study of the Effect of Anatomical Site on Multiple Differentiation of Adipose-Derived Stem Cells in Rats

“…These diets provide energy and protein levels of 359 Kcal/100 g and 23.1 g/100 g, respectively. Euthanasia of the animals was performed with the inhalation method (isoflurane over-dose), according to Siennicka et al [ 31 ].…”

Characterisation of Extracellular Vesicles from Equine Mesenchymal Stem Cells

“…As the regenerative and immunomodulatory capacity of MSCs decreases with increasing donor age, the use of allogeneic MSCs to produce EVs represents an attractive cell-free off-the-shelf treatment option, which allows the selection of optimal donor cells, scalable, standardised manufacturing and treatment optimisation for the target disease [ 79 , 80 ]. However, extensive culture expansion drives MSCs toward replicative senescence and a consequent decline in quality with diminished immunosuppressive and regenerative capacity and proinflammatory features [ 81 , 82 ]. Indeed long-term in vitro culture (high passage) was recently shown to have a greater effect on MSCs (increased β-Galactosidase level) than the natural ageing process of their donor [ 82 ].…”

Heterogeneity of In Vitro Expanded Mesenchymal Stromal Cells and Strategies to Improve Their Therapeutic Actions

“…Several reports have evidenced that MSCs fate is influenced by multiple variables, including the age and pathological condition of the donor [ 13 , 14 ]; tissue source [ 15 ]; culture medium composition [ 16 , 17 ]; passage number [ 13 , 18 , 19 ] or environmental culture conditions, such as oxygen levels, pH or temperature [ 20 , 21 , 22 , 23 ]. All these variables may induce cellular and molecular changes through the culture process that affect the therapeutic potential of MSCs.…”

“…An additional limitation of expanding MSCs is that senescence processes become activated. Those may be even accelerated under specific culture conditions, inducing MSC functional alterations, such as decreased self-renewal, immunomodulation and differentiation [ 13 , 19 ]. Remarkably, the biological features of in vitro senescent MSCs were similar to those found in MSCs obtained from aged donors, including a decreased proliferation ability and differentiation potential [ 13 , 82 ].…”

“…In this context, the use of freshly or refreshed MSCs has been suggested to improve the efficacy in cell-based therapy. The use of low-passaged MSCs is also recommended to achieve better in vitro expansion [ 13 , 19 ].…”