Human T-Cell Isolation

May 12, 2016

Northwestern
Neurological Surgery
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

STEMCELL

Product Name:

Human T cell enrichment kit

Catalog Number:

19051

The kit provided by STEMCELL allows for facilitated isolation of human lymphocytes from peripheral blood mononuclear cells. It involves a magnetic negative selection process which ensures a clean/pure lymphocytic population. Moreover, the process itself is not extremely cytolytic as other kits and provides a great cellular viability result.

Experimental Design and Results Summary

Application

Cell separation

Starting Material

Peripheral Blood Mononuclear Cells

Protocol Overview

Aliquoting 50uL of enrichment solution to 1mL of peripheral blood mononuclear cells in a polystyrene tube and incubating for 10 minutes. Thereafter, adding 50uL of magnetic beads with a 5 minute incubation period. Adding 1mL of required media and placing in magnet for 5 minutes. After the 5 minute incubation, the unwanted cells are stuck to the sides of the tube and the poured out solution is pure with lymphocytes.

Tips

Vortex magnetic beads for 30 seconds prior to addition to cells

Results Summary

>95% lymphocyte viability (assessed with trypan blue)

Additional Notes

None

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Summary

The Good

Easy to use. Can isolate a large number of cells at a time.

The Bad

Expensive depending on frequency of utilization. Have to purchase a magnet separately which may be costly.

The Bottom Line

In order to avoid the prolonged sorting time incurred via the utilization of FACS, this method of lymphocytic isolation is fast/quick/and easy to use.

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