Molecular and cellular characteristics of human and non-human primate multipotent stromal cells from the amnion and bone marrow during long term culture.
Molecular and cellular characteristics of human and non-human primate multipotent stromal cells from the amnion and bone marrow during long term culture.
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DOI：
10.1186/s13287-015-0146-6
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发表时间：
2015-08-22
影响因子：
7.5
通讯作者：
Müller T
中科院分区：
文献类型：
--
作者：
Pogozhykh O;Pogozhykh D;Neehus AL;Hoffmann A;Blasczyk R;Müller T
多能基质细胞（MSC）是再生医学的关键候选细胞之一。然而，MSC 来源的多样性和普遍的异质性导致功能表征数据存在争议。此外，尽管广泛用作临床前动物模型，但人们对普通狨猴的 MSC 知之甚少。对源自人类和普通狨猴的胎盘羊膜和骨髓样本的 MSC 进行 12 代平行表征，通过免疫组织化学、流式细胞术、实时 PCR、代谢活性测试，监测 MSC 标记物和主要组织相容性复合体 (MHC) I 类表达的相似性和显着差异（p ≤ 0.05，学生 t 检验），特别关注多能性相关基因。对人和非人灵长类 MSC 的 MSC 标记表达和分化为间充质谱系的能力进行了表征。 MSC 可以培养超过 100 天（26 代），但与骨髓 MSC 相比，羊膜 MSC 的代谢活性显着增强。有趣的是，在人和狨猴中，羊膜间充质干细胞中 MHC I 类表达显着降低，直至第 6 代，但在骨髓细胞中则不然。对于MSC标志物，羊膜间充质干细胞中的CD73和CD105水平保持不变，并且在传代后期骨髓中略有下降；与狨猴相比，CD166 在人类 MSC 中的表达显着较高，而 CD106 的表达显着较低。所有培养的 MSC 均显示多能性标记物表达，如第 3 代的 Oct-4A，随着时间的推移（第 6-12 代）显着降低，而 Nanog 表达在人骨髓 MSC 中最高。此外，与狨猴相比，人类 MSC 表现出最高的 Sox2 水平，而狨猴则表现出显着更高的 Lin28A 值。 Oct-4 启动子区域的亚硫酸氢盐测序显示，与人类相比，狨猴的 CpG 岛甲基化较少。关于长期培养过程中普通狨猴与人类临床前动物模型的 MSC 特征知之甚少。研究的人类和普通狨猴样本具有许多相似的特征，例如大多数 MSC 标记以及羊膜细胞与骨髓相比 MHC I 类表达减少。此外，多能性标记表明这两个物种的 MSC 亚群具有真正的“干性”，这可以解释它们的高增殖能力，尽管人类和狨猴在 Lin28A 和 Sox2 表达方面存在差异。
Multipotent 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. MSCs 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. Human 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. Little 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.