Abstract
IntroductionMultipotent stromal cells (MSCs) are among the key candidates in regenerative medicine. However variety of MSC sources and general heterogeneity lead to controversial data in functional characterization. Furthermore, despite intensive usage as preclinical animal model, little is known about MSCs of the common marmoset monkey.MethodsMSCs derived from placental amnion and bone marrow samples from human and common marmoset were characterized in parallel over 12 passages to monitor similarities and significant differences (p ≤ 0.05, Student’s t-test) in MSC markers and major histocompatibility complex (MHC) class I expression by immunohistochemistry, flow cytometry, real-time PCR, metabolic activity test, with special focus on pluripotency associated genes.ResultsHuman and non-human primate MSCs were characterized for expression of MSC markers and capability of differentiation into mesenchymal lineages. MSCs could be cultured more than 100 days (26 passages), but metabolic activity was significantly enhanced in amnion vs. bone marrow MSCs. Interestingly, MHC class I expression is significantly reduced in amnion MSCs until passage 6 in human and marmoset, but not in bone marrow cells. For MSC markers, CD73 and CD105 levels remain unchanged in amnion MSCs and slightly decline in bone marrow at late passages; CD166 is significantly higher expressed in human MSCs, CD106 significantly lower vs. marmoset. All cultured MSCs showed pluripotency marker expression like Oct-4A at passage 3 significantly decreasing over time (passages 6–12) while Nanog expression was highest in human bone marrow MSCs. Furthermore, human MSCs demonstrated the highest Sox2 levels vs. marmoset, whereas the marmoset exhibited significantly higher Lin28A values. Bisulfite sequencing of the Oct-4 promoter region displayed fewer methylations of CpG islands in the marmoset vs. human.ConclusionsLittle is known about MSC characteristics from the preclinical animal model common marmoset vs. human during long term culture. Studied human and common marmoset samples share many similar features such as most MSC markers and reduced MHC class I expression in amnion cells vs. bone marrow. Furthermore, pluripotency markers indicate in both species a subpopulation of MSCs with true ‘stemness’, which could explain their high proliferation capacity, though possessing differences between human and marmoset in Lin28A and Sox2 expression.
Highlights
Multipotent stromal cells (MSCs) are among the key candidates in regenerative medicine
There are divergent reports about possible culture duration of MSCs [1, 6, 9] and changes in the expression of MSC markers and major histocompatibility complex (human leukocyte antigen (HLA)/MHC class I, which is a major obstacle in transplantation) over time [6, 10]
The antibodies against human antigens reacted with marmoset samples in immunohistochemical staining displaying CD90+, CD105+, Snail1+, and Bra+, but CD34− (Fig. 1a–j)
Summary
Multipotent stromal cells (MSCs) are among the key candidates in regenerative medicine. Multipotent stromal cells (MSCs) are among the key candidates in the broad perspective of application in the field of regenerative medicine, tissue engineering, and cell replacement therapy. This status is determined by their relative availability from various sources, high plasticity, and immunomodulatory properties. Owing to the variety of MSC sources, as well as the heterogeneity of the derived cell populations in primary cultures, many controversial results exist from different groups in terms of various functions and the general characterization of MSCs [5,6,7]. The expression of pluripotency genes such as Oct-4A, Nanog, Sox, Klf, and c-Myc in MSCs is still discussed controversially [9, 11,12,13,14]
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