The in vitro culture of induced pluripotent stem (iPS) cells or embryonic stem (ES) cells can be a tricky process. Exposing cells to the necessary environmental stimuli to maintain them in an undifferentiated state or conversely direct cell differentiation, can mean juggling cocktails of growth factors, cytokines and inhibitory factors.
Working out what is having the desired effect on your culture is not always easy.
FUJIFILM Wako’s range of recombinant lectin products can aid identification of undifferentiated cells and enable the optimisation of culture conditions to deliver the best results.
The selective binding of recombinant BC2LCN (rBC2LCN) to undifferentiated stem cells was discovered by the Centre for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), Japan1. Following microarray profiling of glycan expression in human iPS and ES cells, rBC2LCN was shown to exclusively recognise an intramolecular sugar chain of podocalyxin found on the surface of undifferentiated human iPS/ES cells2.
Wako’s rBC2LCN has been successfully used numerous studies to identify undifferentiated cell populations via cell staining and flow cytometry protocols once labelled with FITC or Cy33-4. Pre-labelled FITC, 547nm or 635nm fluorochrome probes are now available. Low cytotoxicity means there is no need to wash them out in order to continue culturing cells. If you do need to remove the stain, there is no need to disperse cells, just incubate in rBC2LCN stripping solution for 30 minutes. When proceeding to transplantation models the ability to screen out undifferentiated potentially tumorigenic cells is essential. Recombinant lectin-toxin fusion protein rBC2LCN-PE23 contains the catalytic domain of Pseudomonas aeruginosa exotoxin A and addition of this fusion protein to culture media selectively eliminates any undifferentiated cell populations contaminating the differentiated cell culture.
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See the range of recombinant BC2LCN reagents