I would like to identify adult stem cells by their specific cell markers by way of magnetic bead coulped antibodies coupled to these markers. Is this possible, or do I have to do histo?
Hi Erik,
I think histo is the best option. SInce you probably need multiple markers to define the cells, you best use immunofluorescence.
I can send you some protocols if you want.
With kind regards,
Ronald
Either technology: histology of sectioned tissues or flow of suspended cells works with adult stem cells, we have used both. With respect to antibodies: telomerase-positive adult stem cells: human - CD66e, CD10, CD90, CD13, CD56; non-human - CEA-CAM-1, SSEA-1, SSEA-3, SSEA-4, Thy-1. Telomerase-negative adult "stem" cells - CD105, CD123, CD166, plus others. I work almost exclusively with the telomerase-positive adult stem cells. If you are going to fix your tissues before analysis you need to be cognizant that fixatives can alter antibody binding. I have attached a paper that lists protocols for both techniques as well as optimal fixative to use.
Hi Erik,
Antibody coupled magnetic beads are used to isolate (enrich) the adult stem cells.
To identify the stem cells, you use fluorescent coupled Antibody for Flow cyto analysis on cell suspension, or fluorescent microscopy on sectioned tissue specimen or cytologic specimen (from cell suspension)
You can also use enzyme (horse radish peroxidase or alkaline phosphatase) coupled antibody for sectioned tissue or cytologic specimen.
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