Reprogramming factors involved in hybrids and cybrids of human embryonic stem cells fused with hepatocytes
| Autor: | Linh K. Nguyen, R. Tayfur Tecirlioglu, Alan O Trounson, Karen Koh, Graham Jenkin, Jitong Guo |
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| Rok vydání: | 2010 |
| Předmět: |
Somatic cell
Cellular differentiation Green Fluorescent Proteins Lewis X Antigen Embryoid body Biology Hybrid Cells Transfection Cytoplast Cell Line Cell Fusion Humans reproductive and urinary physiology Embryoid Bodies Embryonic Stem Cells DNA Primers Cell fusion Base Sequence Reverse Transcriptase Polymerase Chain Reaction Cell Differentiation Cell Biology Cell sorting Cell Dedifferentiation equipment and supplies Embryonic stem cell Molecular biology Immunohistochemistry Karyotyping embryonic structures Hepatocytes biological phenomena cell phenomena and immunity Reprogramming Octamer Transcription Factor-3 Developmental Biology Biotechnology |
| Zdroj: | Cellular reprogramming. 12(5) |
| ISSN: | 2152-4998 |
| Popis: | Embryonic stem cells (ESCs) have the potential to reprogram somatic cells into ESC-like cells through cell fusion. In the present study, the potential of human (h)ESC cytoplasts and karyoplasts to reprogram human hepatocytes was evaluated. Green fluorescent protein (GFP) transfected hESCs (ENVY cells) were fused with SNARF-1 (CellTracker)-labeled human hepatocytes using polyethylene glycol (PEG) and fluorescence-activated cell sorting (FACS) to produce hESC-hepatocyte hybrids. Immunocytochemical analysis of ESC markers showed that the hybrids expressed OCT4, TRA-1-60, TRA-1-81, SSEA-4, and GCTM-2. However, SSEA-1, which is typically low or absent on hESCs, was detected on hESC–hepatocyte hybrids. Moreover, reverse transcriptase polymerase chain reaction (RT-PCR) showed that alpha-fetoprotein, which is highly expressed in hepatocytes, was erased in the hybrids. These results indicated that hESCs have the potential to reprogram hepatocyte phenotype to a relatively undifferentiated state, but such hybrid cells are not identical to hESCs. Although hESC–hepatocyte hybrids were aneuploid, they were able to differentiate into embryoid bodies and some types of somatic cells. Furthermore, cybrids of enucleated hESCs and hepatocytes were produced by cell fusion, but the cybrids were unable to self-renew in the same way as hESCs. Presumably, the reprogramming factors are associated with the karyoplast and not the cytoplast of hESCs. |
| Databáze: | OpenAIRE |
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