Ennis J, Götherström C, Le Blanc K, Davies J E
Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
Cytotherapy. 2008;10(2):174-81. doi: 10.1080/14653240801891667.
It has been shown recently that human umbilical cord perivascular cells (HUCPVC) are bio-equivalent to bone marrow-derived mesenchymal stromal cells (BM-MSC) in their mesenchymal differentiation and marker expression. HUCPVC populations provide high yields of rapidly proliferating mesenchymal progenitor cells. The question we wished to address, in two independent laboratory studies, was whether HUCPVC exhibit a similar in vitro immunologic phenotype to that of BM-MSC.
HUCPVC were isolated by physical extraction of umbilical vessels followed by enzymatic digestion of the perivascular cells, and lymphocytes were obtained from heparinized human peripheral blood. Experimental evaluations were lymphocyte proliferation in HUPCVC or BM-MSC co-cultures with peripheral blood lymphocytes (PBL), mixed lymphocyte cultures (MLC) containing BM-MSC or HUCPVC, CD25 and CD45 expression in co-cultures containing HUCPVC, and finally lymphocyte proliferation in TransWell MLC with HUCPVC.
Both HUCPVC and BM-MSC showed no significant increase in proliferation of lymphocytes when co-cultured. The addition of 10% HUCPVC or BM-MSC significantly reduced proliferation of PBL in one-way MLC. Upon inclusion of HUCPVC with activated T-cell lines, the expression of both CD25 and CD45 showed a significant decrease. HUCPVC were able to reduce lymphocyte cell numbers significantly when separated with a membrane insert.
HUCPVC are not alloreactive and exhibit immunosuppression in vitro. Lymphocyte activation is significantly reduced in the presence of HUCPVC, and the immunosuppressive effect of HUCPVC is due, in part, to a soluble factor. Thus HUCPVC shows a similar immunologic phenotype to BM-MSC.
最近的研究表明,人脐带血管周围细胞(HUCPVC)在间充质分化和标志物表达方面与骨髓来源的间充质基质细胞(BM-MSC)具有生物等效性。HUCPVC群体可提供高产量的快速增殖间充质祖细胞。我们希望在两项独立的实验室研究中解决的问题是,HUCPVC在体外是否表现出与BM-MSC相似的免疫表型。
通过物理提取脐带血管,然后酶消化血管周围细胞来分离HUCPVC,从肝素化的人外周血中获取淋巴细胞。实验评估包括HUPCVC或BM-MSC与外周血淋巴细胞(PBL)共培养中的淋巴细胞增殖、含有BM-MSC或HUCPVC的混合淋巴细胞培养(MLC)、含有HUCPVC的共培养中CD25和CD45的表达,最后是HUCPVC在TransWell MLC中的淋巴细胞增殖。
HUCPVC和BM-MSC在共培养时淋巴细胞增殖均未显著增加。在单向MLC中添加10%的HUCPVC或BM-MSC可显著降低PBL的增殖。当将HUCPVC与活化的T细胞系一起培养时,CD25和CD45的表达均显著降低。当用膜插入物分隔时,HUCPVC能够显著减少淋巴细胞数量。
HUCPVC无同种异体反应性,在体外表现出免疫抑制作用。在有HUCPVC存在时,淋巴细胞活化显著降低,HUCPVC的免疫抑制作用部分归因于一种可溶性因子。因此,HUCPVC表现出与BM-MSC相似的免疫表型。