Related Experiment Video
Updated: Aug 8, 2026
Efficient Derivation of Human Neuronal Progenitors and Neurons from Pluripotent Human Embryonic Stem Cells with Small Molecule Induction
Published on: October 28, 2011
Cloned human teratoma cells differentiate into neuron-like cells and other cell types in retinoic acid
Abstract:
Single cell clones were isolated from the human teratoma line, Tera-2. The cells of three of these clones were studied. The progressively growing cells were shown to be tumorigenic, and they were characterized by the lack of expression of beta 2-microglobulin and HLA-A,B,C determinants on the cell surface. The majority of the cells expressed Thy-1 antigen and a 90 X 10(3) molecular weight protein recognized by the monoclonal antibody F10.44.2; between a third and half of the cells expressed the sugar specificities detected by the anti-SSEA-1 monoclonal antibody. In response to 5 X 10(-5) M-retinoic acid applied to cells in monolayer culture, the cells differentiated into a population of flat static cells arrested in the G1 phase of the cell cycle. A substantial proportion of these differentiated cells expressed beta 2-microglobulin and 43 X 10(3) molecular weight HLA-A,B,C polypeptides, Thy-1, SSEA-1 sugar determinants, and the 90 X 10(3) Mr protein recognized by F10.44.2. The apparent molecular weight of fibronectin secreted by the cells decreased by about 5 X 10(3) Mr to 235 X 10(3) Mr after differentiation. The progressively growing cells lacked reactivity with reagents that mark cells in the nervous system. Following aggregation and retinoic acid treatment, neuron-like cells were formed. These cells reacted with reagents that also react with human neurons in culture: they reacted with tetanus toxin, the anti-neurofilament antibodies BF10 and RT97, the anti-ganglioside, GQ1c antibody F12 A2B5, and anti-Thy-1. The progressively growing cells of these Tera-2 clones are therefore capable of forming at least two types of cell: the flat cells in monolayer cultures and the neuron-like cells. None of the cell populations reacted with the monoclonal antibody against SSEA-3 and these cloned cells are therefore distinct from previous isolates from Tera-2.
Insights
Human teratoma cells (Tera-2 clones) can differentiate into flat cells or neuron-like cells when treated with retinoic acid, altering their surface markers and cell cycle. These findings reveal the developmental potential of Tera-2 clones.
Area of Science:
- Developmental Biology
- Cancer Research
- Cell Biology
Background:
- Human teratoma cell lines, such as Tera-2, are valuable models for studying cellular differentiation and tumorigenesis.
- Understanding the molecular and phenotypic changes during differentiation is crucial for cancer research and regenerative medicine.
- Previous studies have identified specific markers and responses to differentiation agents in teratoma cells.
Purpose of the Study:
- To investigate the differentiation potential of single cell clones isolated from the Tera-2 human teratoma line.
- To characterize the phenotypic changes, including cell surface marker expression and cell cycle status, upon retinoic acid-induced differentiation.
- To determine if these clones can differentiate into neural lineages.
Main Methods:
- Isolation and propagation of single cell clones from the Tera-2 teratoma line.
- Tumorigenicity assessment of progressively growing cells.
- Characterization of cell surface markers using monoclonal antibodies (e.g., anti-beta 2-microglobulin, anti-HLA-A,B,C, anti-Thy-1, anti-SSEA-1).
- Induction of differentiation using retinoic acid in monolayer cultures and aggregated cultures.
- Analysis of cell cycle progression (G1 arrest).
- Assessment of fibronectin secretion.
- Immunological characterization of differentiated cells, including neuron-specific markers (e.g., neurofilaments, gangliosides).
Main Results:
- Progressively growing Tera-2 clones were tumorigenic and lacked beta 2-microglobulin and HLA-A,B,C surface expression.
- Retinoic acid treatment induced differentiation into flat cells arrested in G1 phase, with altered expression of beta 2-microglobulin, HLA-A,B,C, Thy-1, SSEA-1, and a reduction in fibronectin molecular weight.
- Aggregation and retinoic acid treatment led to the formation of neuron-like cells expressing neural markers (tetanus toxin, neurofilament antibodies, anti-ganglioside antibody, anti-Thy-1), distinct from previous Tera-2 isolates (negative for SSEA-3).
Conclusions:
- The studied Tera-2 clones possess the capacity to differentiate into at least two distinct cell types: flat cells and neuron-like cells.
- Retinoic acid is an effective inducer of differentiation in these Tera-2 clones, leading to significant phenotypic and molecular changes.
- These findings highlight the multipotent differentiation potential of specific Tera-2 clones and their utility in studying early human cell differentiation and neural development.
