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A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell
K Kobayashi1, T Ohye, I Pastan
1Institute for Comprehensive Medical Science, School of Medicine, Fujita Health University, Aichi, Japan.
Abstract:
Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor alpha subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation.
Insights
Immunotoxoin-mediated cell targeting (IMCT) effectively eliminated specific mouse embryonic stem (ES) cells expressing a targeted receptor. This demonstrates IMCT
Area of Science:
- Stem cell biology
- Molecular biology
- Immunology
Background:
- Immunotoxoin-mediated cell targeting (IMCT) uses recombinant immunotoxins to eliminate specific cell types.
- Mouse embryonic stem (ES) cells are crucial for developmental biology research and therapeutic applications.
Purpose of the Study:
- To evaluate the feasibility of IMCT for negative selection in mouse ES cells.
- To assess the efficacy of anti-Tac(Fv)-PE40 in targeting ES cells expressing the human interleukin-2 receptor alpha subunit.
Main Methods:
- Utilized recombinant immunotoxin anti-Tac(Fv)-PE40 for targeted cell ablation.
- Investigated the responsiveness of mouse ES cells engineered to express the human interleukin-2 receptor alpha subunit.
Main Results:
- The immunotoxin treatment selectively eliminated ES cells that expressed the target receptor.
- Confirmed the target specificity of anti-Tac(Fv)-PE40 in the context of mouse ES cells.
Conclusions:
- IMCT is a viable strategy for negative selection of specific cell populations in mouse ES cells.
- This technique can be employed for enriching ES cell clones with desired genetic modifications.