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Isolation and partial characterization of lectin-resistant F9 cells
Abstract:
Cytotoxic plant lectins have been used for the single-step selection of mouse embryonal carcinoma cell mutants with altered expression of surface glycoconjugates. Following mutagenesis, several F9 and OTF9-63 cell lines resistant to the lectins from Triticum vulgaris or Ricinus communis were obtained. At least five distinct lectin-resistant (LecR) phenotypes have been identified on the basis of their relative sensitivities to four different plant lectins and their altered lectin-binding properties. None of the mutant types exhibits a significant change in the ability to bind a monoclonal antibody against the stage-specific embryonic antigen, SSEA-1. All of the mutants form aggregates when cultured in bacteriological petri dishes and appear to differentiate into endoderm-like cells following exposure to retinoic acid. However, two of the LecR cell lines exhibit an altered morphology when grown on a plastic substratum.
Insights
Researchers developed cytotoxic plant lectin-resistant (LecR) cell lines from mouse embryonal carcinoma cells. These LecR mutants exhibit altered surface glycoconjugates and distinct differentiation responses, offering insights into cell surface molecule functions.
Area of Science:
- Cell Biology
- Developmental Biology
- Biochemistry
Background:
- Cytotoxic plant lectins are valuable tools for selecting cell mutants.
- Altered surface glycoconjugates can impact cell behavior and differentiation.
Purpose of the Study:
- To generate and characterize mouse embryonal carcinoma cell mutants resistant to cytotoxic plant lectins.
- To investigate the functional consequences of altered surface glycoconjugate expression.
Main Methods:
- Mutagenesis of F9 and OTF9-63 cell lines.
- Selection using Triticum vulgaris and Ricinus communis lectins.
- Phenotypic characterization of lectin-resistant (LecR) mutants using multiple lectins and antibody binding assays.
Main Results:
- Several LecR cell lines were obtained, exhibiting at least five distinct phenotypes based on lectin sensitivity and binding.
- Mutants showed no significant change in binding to anti-SSEA-1 antibody.
- All mutants formed aggregates and differentiated into endoderm-like cells upon retinoic acid exposure.
- Two LecR lines displayed altered morphology on plastic substrata.
Conclusions:
- Lectin resistance provides a method to isolate cell mutants with altered surface glycoconjugates.
- These mutants retain some differentiation capabilities but may exhibit changes in cell morphology.
- The study highlights the role of specific surface glycoconjugates in cell behavior and differentiation.