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Sterile Tissue Harvest

Overview

Source: Kay Stewart, RVT, RLATG, CMAR; Valerie A. Schroeder, RVT, RLATG. University of Notre Dame, IN

In 1959 The 3 R's were introduced by W.M.S. Russell and R.L. Burch in their book The Principles of Humane Experimental Technique. The 3 R's are replacement, reduction, and refinement of the use of animals in research.¹ The use of cell lines and tissue cultures that originated from research animals is a replacement technique, as it allows for many experiments to be conducted in vitro. Harvesting tissues and organs for use in cell and tissue cultures requires aseptic technique to avoid contamination of the tissues. Sterile harvest is also necessary for protein and RNA analysis and metabolic profiling of tissues. This manuscript will discuss the process of sterile organ harvest in rats and mice.

Procedure

1. Preparation

Materials commonly used include 70% alcohol in a squirt bottle, sterile 20-25 gauge needles, two packs of sterile instruments of iris scissors and thumb forceps (one for the skin and one for the internal tissues), paper towels, sterile surgeon gloves, a sterile petri dish, sterile medium, and sterile saline.
The hood fan should be running for at least 15 minutes prior to the procedure, and it should be disinfected with 70% alcohol.
If the hood is equipped with a UV steriliza

Application and Summary

Although many of the procedures for sterile organ harvest are similar to a standard necropsy, the use of sterile technique is imperative for tissues to be of value for experimental procedures, which include harvesting cells from the excised tissues. Profiling or collecting RNA from tissues can be compromised if sterility is not maintained.

References

Russel, W.M.S. and Burch, R.L. 1959. The Principles of Humane Experimental Technique. Methuen: London.
Petucci, C., Rojas-Betancourt, S., and Gardell, S.J. 2012. Comparison of tissue harvest protocols for the quantitation of acylcarnitines in mouse heart and liver by mass spectrometry. Metabolomics. 8.784-792.
Parkinson, C.M., O'Brien, A., Albers, T.M., Simon, M.A., Clifford, C.B., and Pritchett-Corning, K.R. 2011. Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice. 54. e2966. 1-7.