Effects of Donor Age, Long-Term Passage Culture, and Cryopreservation on Tonsil-Derived Mesenchymal Stem Cells

Abstract

Objectives: Human mesenchymal stem cells (MSCs) are efficacious in various cellular therapeutic applications and have been isolated from several tissues. Recent studies have reported that human tonsil tissue contains a new source of progenitor cells, potentially applicable for cell-based therapies. Information about the effects of donor age, long-term passage and cryopreservation are essential for clinical applications and cell-based therapies. Therefore, the authors investigated how the morphology, cell-surface markers, proliferation potential and differentiation capacity of tonsil-derived MSCs (T-MSCs) were affected by donor age, long-term passage, and cryopreservation. Materials and Methods: T-MSCs were isolated from tonsillar tissue of 20 patients undergoing tonsillectomy. Authors evaluated the effects of donor-age, long-term passage, and cryopreservation on the morphology, surface markers, proliferation potential and differentiation capacities of T-MSCs. Results: T-MSCs exhibited a fibroblast-like, spindle-shaped appearance. There were no significant morphological differences according to donor age, long-term passage or cryopreservation. T-MSCs isolated from donors of various ages were positive for markers CD90, CD44, and CD73, but negative for CD45, CD31, and HLA-DR. There were no significant differences in the expression of positive and negative surface markers as a function of donor age, long-term passage and cryopreservation. T-MSCs from different donor age groups showed similar proliferation potentials after passage 2. After long-term passage and cryopreservation, there were no significant morphological differences. Cryopreservation did not affect the proliferation potential of T-MSCs, but there was a significant decrease in the proliferation potential in long-term passage T-MSCs (passage 15). The effect of donor age, long-term passage and cryopreservation on the in vitro adipogenic, osteogenic, and chondrogenic differentiation potential of T-MSCs was not significant. Conclusion: The effect of donor age, long-term passage culture, and cryopreservation on T-MSC properties are negligible, except for the proliferation capacity of long-term cultured T-MSCs. Therefore, T-MSCs are considered to be promising MSCs that can be used as future alternative sources for autologous or allogenic MSCs.

Differentiation of Human Tonsil-Derived Mesenchymal Stem Cells into Schwann-Like Cells Improves Neuromuscular Function in a Mouse Model of Charcot-Marie-Tooth Disease Type 1A

“…As tonsillar tissues are discarded after surgery, the isolation of stem cells from these discarded tissues offers another valuable means of recycling human tissue for stem cell therapy [20,23]. In addition, the T-MSCs have MSC characteristics and are known to secrete various cytokines [27], growth factors [28], and exosomes [29] considered suitable for cell therapy.…”

“…In addition, the T-MSCs have MSC characteristics and are known to secrete various cytokines [27], growth factors [28], and exosomes [29] considered suitable for cell therapy. The T-MSCs show excellent immune modulation activity and no immunologic problems even when the T-MSCs were combined from three independent donors [23]. Thus, we speculated that the T-MSCs would be suitable for clinical use as a heterologous transplantation cell source.…”

“…T-MSCs are useful for stem-cell therapy in various disease conditions because tonsils are a ready source of stem cells [23,24]. T-MSCs have the potential to differentiate into Schwann-like cells and can secrete neurotrophic factors to promote axonal growth and remyelination [18].…”

Multiple Directional Differentiation Difference of Neonatal Rat Fibroblasts from Six Organs

“…It was widely believed that mesenchymal stem cells (MSCs) were multipotential fibroblast-like cells with CD73+, CD105+, CD14-, CD34-and CD45-and plastic adherent [25][26][27]. Recent studies had revealed that fibroblasts shared much more in common than previously recognized.…”

Isolation and Localization of Mesenchymal Stem Cells in Human Palatine Tonsil by W5C5 (SUSD2)

“…Recent studies reported the isolation of tonsilMSCs (T-MSCs) from tonsil tissues and found that the morphology, cell-surface markers, proliferation potential, and differentiation potential of T-MSCs were similar to those of BMMSCs [11,15]. T-MSCs were considered to be promising MSCs that could be used as future alternative sources for autologous or allogenic MSCs, because the effects of donor age, longterm passage culture, and cryopreservation on T-MSC properties were negligible [11,15]. Despite the reports of T-MSCs as a new autologous or homologous source of MSCs, there have been no reports on specific markers of MSCs for purified T-MSC isolation.…”