N-Glycolylneuraminic Acid Xenoantigen Contamination of Human Embryonic and Mesenchymal Stem Cells Is Substantially Reversible

Abstract

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. STEM CELLS 2007;25: 197-202

How has dental pulp stem cells isolation been conducted? A scoping review

“…These processes could influence the survival and efficacy of the transplanted cells in clinical applications. 35,36 Thus, alternative supplements have been investigated. Serum-free media have been continuously tested for stem cell culture; however, so far, the literature indicates that these media do not adequately support DPSCs proliferation and differentiation.…”

Influence of culture conditions on expansion and re-differentiation of chondrocytes from horses of different ages

“…However, poor chondrocyte attachment in autologous sera at each passage resulted in lower final cell yields compared to the other sera tested. Development of serum free or autologous serum culturing systems for cell expansion in tissue engineering aims to circumvent the problem of disease transmission and immunogenic reaction (Heng et al 2004, Heiskanen et al 2007). Several studies have demonstrated beneficial effect of growth factor addition to the serum-free expansion medium for the proliferation and maintenance of the chondrogenic phenotype (Mandl et al 2002, Malpeli et al 2004, even overcoming the age-related differences (Giannoni et al 2005).…”

“…The exposure of cells to FCS however, has several disadvantages including possible viral or bacterial infections (Dedrick 1997) and transmission of prions (Klein and Dumble 1993). If the cells have been exposed to FCS during cultivation, the following transplantation may induce immune or local inflammatory responses (Selvaggi et al 1997, Heiskanen et al 2007. Therefore, the development of FCS-free culturing protocols certainly represents an advantage for future development of cellular therapies, and the use of serum-free media, allogenic, and autologous serum have being already investigated as potential alternatives for growing cells in human regenerative medicine (Stute et al 2004, Shahdadfar et al 2005, Tallheden et al 2005.…”

A Proliferation and surface marker characterization of adipose stem cells after culture in various processed outdated platelet lysate containing media

“…7 Moreover, FBS recently are identified to contain xeno-protein that could not be removed, i.e. N-Glycolylneuraminic acid (Neu5Gc), 8,9 which might be harmful if the stem cells are intended to be used as therapy in human.…”