Intrinsic properties and external factors determine the differentiation bias of human embryonic stem cell lines
| Autor: | Ashish Mehta, Sen Anish Majumdar, Chandra Viswanathan, Sheena Mathew |
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| Rok vydání: | 2010 |
| Předmět: |
Homeobox protein NANOG
Pluripotent Stem Cells Cellular differentiation Cell Culture Techniques Down-Regulation Fluorescent Antibody Technique Gene Expression Germ layer Biology Cell Line Mesoderm Downregulation and upregulation Ectoderm medicine Humans Cell Lineage Embryoid Bodies Embryonic Stem Cells Genetics Homeodomain Proteins Reverse Transcriptase Polymerase Chain Reaction Mesenchymal stem cell Gene Expression Regulation Developmental Cell Differentiation Cell Biology General Medicine Nanog Homeobox Protein Flow Cytometry Embryonic stem cell Cell biology Up-Regulation medicine.anatomical_structure Cell culture embryonic structures Endoderm Octamer Transcription Factor-3 Biomarkers |
| Zdroj: | Cell biology international. 34(10) |
| ISSN: | 1095-8355 |
| Popis: | A major goal of human embryonic stem cell (hESC) research is to regulate differentiation through external means to generate specific cell types with high purity for regenerative medicine applications. Although all hESC lines express pluripotency-associated genes, their differentiation ability to various lineages differs considerably. We have compared spontaneous differentiation propensity of the two hESC lines, RelicellhES1 and BG01. Spontaneous differentiation of hESC lines grown in different media conditions was followed by differentiation using two methods. Kinetic data generated by real-time gene expression studies for differentiated cell types were analyzed, and confirmed at protein levels. Both cell lines showed upregulation of genes associated with the 3 germ layers, although stark contrast was evident in the magnitude of upregulation of lineage specific genes. A distinct difference was also found in the rate at which the pluripoteny factors, Oct-4 and Nanog, were downregulated during differentiation. Once differentiation was initiated, both Oct-4 and Nanog gene expression was drastically reduced in RelicellhES1, whereas a gradual decrease was observed in BG01. A clear trend is seen in RelicellhES1 to differentiate into neuroectodermal and mesenchymal lineages, whereas BG01 cells are more prone to mesoderm and endoderm development. In addition, suspension versus plated methods of cell culture significantly influenced the outcome of differentiation of certain types of cells. Results obtained by spontaneous differentiation of hESCs were also amplified by induced differentiation. Thus, differential rates of downregulation of pluripotency markers along with culture conditions seem to play an important role in determining the developmental bias of human ES cell lines. |
| Databáze: | OpenAIRE |
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