Trivedi Parul, Hematti Peiman
Department of Medicine, University of Wisconsin-Madison, School of Medicine and Public Health, Madison, WI 53792-5156, USA.
Exp Hematol. 2008 Mar;36(3):350-9. doi: 10.1016/j.exphem.2007.10.007. Epub 2008 Jan 7.
We have previously shown the simultaneous generation of CD73(+) mesenchymal stromal cells (MSCs) along with CD34(+) hematopoietic cells from human embryonic stem cells (ESCs) when they are cocultured with OP9 murine stromal cells. We investigated whether MSCs can be derived from human ESCs without coculturing with OP9 cells, and if such cells exhibit immunological properties similar to MSCs derived from adult human bone marrow (BM).
Our starting populations were undifferentiated human ESCs cultured on Matrigel-coated plates without feeder cells. The differentiated fibroblast-looking cells were tested for expression of MSC markers and their potential for multilineage differentiation. We investigated surface expression of human leukocyte antigen (HLA) molecules on these MSCs before and after treatment with interferon-gamma (IFN-gamma). We also tested the proliferative response of T-lymphocytes toward MSCs and the effects of MSCs in mixed lymphocyte reaction (MLR) assays.
We derived populations of MSCs from human ESCs with morphology, cell surface marker characteristics, and differentiation potential similar to adult BM-derived MSCs. Similar to BM-derived MSCs, human ESC-derived MSCs express cell surface HLA class I (HLA-ABC) but not HLA class II (HLA-DR) molecules. However, stimulation with IFN-gamma induced the expression of HLD-DR molecules. Human ESC-derived MSCs did not induce proliferation of T-lymphocytes when cocultured with peripheral blood mononuclear cells. Furthermore, ESC-derived MSCs suppressed proliferation of responder T-lymphocytes in MLR assays.
MSCs can be derived from human ESCs without feeder cells. These human ESC-derived MSCs have cell surface markers, differentiation potentials, and immunological properties in vitro that are similar to adult BM-derived MSCs.
我们之前已经表明，当人类胚胎干细胞（ESC）与OP9小鼠基质细胞共培养时，可同时产生CD73(+)间充质基质细胞（MSC）和CD34(+)造血细胞。我们研究了是否可以在不与OP9细胞共培养的情况下从人类ESC中获得MSC，以及这些细胞是否表现出与源自成人骨髓（BM）的MSC相似的免疫特性。
我们的起始细胞群体是在无饲养细胞的基质胶包被板上培养的未分化人类ESC。对分化后的成纤维样细胞进行MSC标志物表达及其多向分化潜能的检测。我们研究了用干扰素-γ（IFN-γ）处理前后这些MSC上人类白细胞抗原（HLA）分子的表面表达。我们还检测了T淋巴细胞对MSC的增殖反应以及MSC在混合淋巴细胞反应（MLR）试验中的作用。
我们从人类ESC中获得了MSC群体，其形态、细胞表面标志物特征和分化潜能与成人BM来源的MSC相似。与BM来源的MSC相似，人类ESC来源的MSC表达细胞表面I类HLA（HLA-ABC）但不表达II类HLA（HLA-DR）分子。然而，IFN-γ刺激可诱导HLD-DR分子的表达。当与外周血单个核细胞共培养时，人类ESC来源的MSC不会诱导T淋巴细胞增殖。此外，ESC来源的MSC在MLR试验中抑制反应性T淋巴细胞的增殖。
无需饲养细胞即可从人类ESC中获得MSC。这些人类ESC来源的MSC在体外具有与成人BM来源的MSC相似的细胞表面标志物、分化潜能和免疫特性。
