C Tubes & gentleMACS Dissociator for Tumor Dissociation

September 14, 2015

University of North Carolina at Chapel Hill
Research
Laboratory Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Miltenyi Biotech

Product Name:

C Tubes & gentleMACS™ Dissociator

Catalog Number:

130-093-237 / 130-093-235

Miltenyi Biotech's C-Tubes are designed to streamline tissue dissociation, and are very helpful in doing so. By using set programs on the accompanying dissociator machine, you can select for specific cell types that you wish to isolate. I used these to dissociate tumor tissue in a cell line derivation procedure and received consistent, thoroughly positive results with my final cell lines.

Experimental Design and Results Summary

Application

Tissue Dissociation / Cell Line Derivation

Starting Material

Mouse Lung Tumor Tissue

Protocol Overview

Once I excised the tumors from the mice, I minced each for several minutes with scalpels under sterile conditions in 5 mL of media and then transferred the minced tumor tissue solution to a C-tube. I then ran the m_lung_01 program 2 times per C-tube on the gentleMACS dissociator. Following a 30 minute incubation with collagenase/dispase, I then ran the m_lung_02 program once per tube on the gentleMACS dissociator. After quenching the collagenase/dispase with EDTA, I put each solution through a 40uM cell strainer, pelleted the resulting single cells with 200g for 3 minutes, resuspended the single-cell solution with media, and plated the resulting cells. Change media 24-48 hours following the initial plating.

Tips

Remember to use the septum-sealed opening to add reagents to the C-tube instead of having to open it and compromise sterility.

Results Summary

Consistent dissociation of lung tumor tissue was seen across more than 5 samples. My target tumor cells survived, dissociated, and attached very well, despite the inside of the tube becoming quite frothy during dissociation. Dissociation of the tumor tissue was thorough enough to render tumor chunks into solution, but not so rough as to kill my target cells. Very few tumor chunks were left on the large side of the cell strainer, and a great deal of cells made it into single-cell solution.

Additional Notes

Make sure you also have a gentleMACS dissociator machine, or these tubes are useless.

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Summary

The Good

Easy to maintain sterility while thoroughly and consistently dissociating tissue

The Bad

Can be expensive, requires a gentleMACS dissociator as well.

The Bottom Line

These tubes are my preferred method of dissociating tissue in cell line derivation procedures that start with solid tissue. The ease of use is top notch, and the variety of programs on the accompanying gentleMACS Dissociator machine allow you to fine-tune the amount of shearing forces required to dissociate specific cell types. I've used these tubes probably 10 times, and have had great results each time.

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