Abstract
Abstract Background: Human embryonic stem (hES) cells are pluripotent, and can differentiate into three germ layers. Traditionally, cultures of hES cells are maintained in a system containing mouse embryonic fibroblasts as a feeder layer for support of undifferentiated growth. However, contamination by animal cells limits the use of hES cells. Objective: We evaluated the use of human dental pulp stem cells (hDPSCs) as a feeder layer for hES cell culture. It should be possible to obtain a new source of human mesenchymal stem cells for feeder cells to maintain undifferentiated growth of hES cells. Methods: hDPSCs from removed impacted wisdom teeth (third molars) were extracted, cultured, and characterized for mesenchymal stem cell properties. Furthermore, hDPSCs were used as a feeder layer for culturing Chula2 and Chula5 hES cell lines. Finally, hES cell lines grown on hDPSCs feeders were examined embryonic stem cell properties. Results: We found that hDPSCs, which have mesenchymal properties, can support undifferentiated growth of hES cell lines. After prolonged culture (passage 17), these hES cell lines still maintain ES cell properties including typical morphology seen in hES cells, the expression of pluripotency markers (Oct4, Sox2, Nanog, Rex1, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), embryoid body formation and retention of a normal karyotype. Conclusion: hDPSCs, derived from the pulp tissue of impacted third molars, are a potential source of human feeder cells for the culture of undifferentiated hES cells.
Highlights
Human embryonic stem cells are pluripotent, and can differentiate into three germ layers
Results human dental pulp stem cells (hDPSCs) expressed fibroblast-like morphology and protein markers of mesenchymal stem cells and differentiated into adipocytes, osteocytes and chondrocytes
HDPSCs expressed mesenchymal protein markers including CD29, CD44, CD73, CD90, and HLA-Dr, but other protein markers such as CD11b, CD34, D45, CD79a, and HLA-ABC were not expressed in these cells (Figure 1c)
Summary
Human embryonic stem (hES) cells are pluripotent, and can differentiate into three germ layers. Cultures of hES cells are maintained in a system containing mouse embryonic fibroblasts as a feeder layer for support of undifferentiated growth. Objective: We evaluated the use of human dental pulp stem cells (hDPSCs) as a feeder layer for hES cell culture. It should be possible to obtain a new source of human mesenchymal stem cells for feeder cells to maintain undifferentiated growth of hES cells. HDPSCs were used as a feeder layer for culturing Chula and Chula hES cell lines. HES cell lines grown on hDPSCs feeders were examined embryonic stem cell properties. Results: We found that hDPSCs, which have mesenchymal properties, can support undifferentiated growth of hES cell lines. Conclusion: hDPSCs, derived from the pulp tissue of impacted third molars, are a potential source of human feeder cells for the culture of undifferentiated hES cells
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