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Updated: Jul 6, 2026

Isolation and Activation of Murine Lymphocytes
Published on: October 30, 2016
Development of a storage-compatible microtiter plate-based technique for lymphocyte proliferation
R P Kerr1, S Kim, S Ansar Ahmed
1The Center for Molecular Medicine and Infectious Diseases, Virginia Tech, Blacksburg, VA 24060-0342, USA.
Insights
A new method for preparing lymphocyte proliferation assay (LPA) plates by coating with complete media and storing at -40°C for up to 10 days yields results comparable to fresh preparations, saving researchers time and labor.
Area of Science:
- Immunology
- Cellular Biology
- Biotechnology
Background:
- The lymphocyte proliferation assay (LPA) is a critical in vitro test for assessing non-specific lymphocyte function.
- Current LPA protocols are time-consuming, labor-intensive, and require fresh reagent preparation, posing challenges for researchers.
- A stable, storage-compatible plate coating technique for LPA could significantly benefit both basic and clinical research.
Purpose of the Study:
- To develop and evaluate a novel, stable, and storage-compatible plate coating technique for lymphocyte proliferation assays.
- To compare the efficacy of different mitogen plate coating methods with traditional fresh preparations.
Main Methods:
- Investigated various mitogen plate coating techniques for lymphocyte proliferation.
- Compared the proliferation response of lymphocytes cultured on pre-coated, stored plates versus freshly prepared plates.
- Plates were coated with complete media and stored at -40°C for up to 10 days.
Main Results:
- Plate coating with complete media and subsequent storage at -40°C for up to 10 days demonstrated comparable results to freshly prepared control plates.
- This storage-compatible method maintained lymphocyte proliferation response without significant loss of function.
Conclusions:
- A stable and storage-compatible plate coating technique for lymphocyte proliferation assays has been successfully developed.
- This method offers a practical solution for researchers, reducing preparation time and labor while maintaining assay reliability.
- The findings support the adoption of this technique for routine use in immunological research and diagnostics.
Abstract:
The lymphocyte proliferation assay (LPA) is an important tier 1 test to assess non-specific lymphocyte function, in vitro. However, this assay requires fresh preparation, is time consuming, and labor intensive. Developing a plate coating technique for lymphocyte proliferation that is both stable and storage compatible would be useful to the basic and clinical researcher. In this study, we compared the effects of different mitogen plate coating techniques on lymphocyte proliferation to freshly prepared plates. The results show that plates prepared with complete media and stored at -40 degrees C up to 10 days corresponded well to control plates.
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