N-glycolylneuraminic acid xenoantigen contamination of human embryonic and mesenchymal stem cells is substantially reversible
N-glycolylneuraminic acid xenoantigen contamination of human embryonic and mesenchymal stem cells is substantially reversible
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DOI：
10.1634/stemcells.2006-0444
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发表时间：
2007-01-01
期刊：
影响因子：
5.2
通讯作者：
Laine, Jarmo
中科院分区：
文献类型：
--
作者：
Heiskanen, Annamari;Satomaa, Tero;Laine, Jarmo
人类胚胎干细胞和间充质干细胞疗法可以为大量患者提供显著益处。然而，最近，据报道，在小鼠饲养细胞上培养的人胚胎干细胞系被异种碳水化合物N-羟乙酰神经氨酸（Neu 5Gc）污染，并被认为可能不适合人类治疗。为了确定Neu 5Gc污染问题的程度，以开发干细胞疗法，我们研究了它是否也发生在人类饲养细胞和间充质干细胞上培养的细胞中，污染的来源是什么，以及污染是否可逆。我们发现，N-羟乙酰神经氨酸存在于人类饲养细胞培养的胚胎干细胞中，与Neu 5Gc的商业血清替代培养基的成分中的存在相关。在胎牛血清存在下培养的间充质干细胞中也发生了类似的污染。结果表明，Neu 5Gc是存在于糖蛋白和脂质连接的聚糖，分别通过质谱分析和单克隆抗体染色检测。值得注意的是，在两种细胞类型的后代中，污染在很大程度上是可逆的，这表明净化的细胞可能来自现有的干细胞系。虽然在暴露于胎牛血清的间充质干细胞的临床试验中尚未报告重大并发症，但免疫原性污染可能反映在移植细胞的活力和功效中，从而使已发表的结果产生偏差。干细胞安全培养条件的定义对于细胞疗法的未来发展至关重要。
Human embryonic and mesenchymal stem cell therapies may offer significant benefit to a large number of patients. Recently, however, human embryonic stem cell lines cultured on mouse feeder cells were reported to be contaminated by the xeno-carbohydrate N-glycolylneuraminic acid (Neu5Gc) and considered potentially unfit for human therapy. To determine the extent of the problem of Neu5Gc contamination for the development of stem cell therapies, we investigated whether it also occurs in cells cultured on human feeder cells and in mesenchymal stem cells, what are the sources of contamination, and whether the contamination is reversible. We found that N-glycolylneuraminic acid was present in embryonic stem cells cultured on human feeder cells, correlating with the presence of Neu5Gc in components of the commercial serum replacement culture medium. Similar contamination occurred in mesenchymal stem cells cultured in the presence of fetal bovine serum. The results suggest that the Neu5Gc is present in both glycoprotein and lipid-linked glycans, as detected by mass spectrometric analysis and monoclonal antibody staining, respectively. Significantly, the contamination was largely reversible in the progeny of both cell types, suggesting that decontaminated cells may be derived from existing stem cell lines. Although major complications have not been reported in the clinical trials with mesenchymal stem cells exposed to fetal bovine serum, the immunogenic contamination may potentially be reflected in the viability and efficacy of the transplanted cells and thus bias the published results. Definition of safe culture conditions for stem cells is essential for future development of cellular therapies.