A simple method for unbiased quantitation of adoptively transferred cells in solid tissues

Area of Science:

• Immunology
• Cell Biology
• Microscopy

Background:

• Accurate quantification of adoptively transferred cells is crucial for immunological studies.
• Existing methods may lack precision or introduce bias in cell enumeration.
• Understanding cell distribution over time is essential for evaluating immune responses.

Purpose of the Study:

• To develop and validate a direct, unbiased method for estimating the total number of adoptively transferred cells in various organs.
• To quantify the organ distribution and kinetics of CFSE-labeled splenocytes over time in a mouse model.
• To provide a precise and straightforward technique for quantitative immunological research.

Main Methods:

• Utilized a mouse model (C57BL/6J) for adoptive cell transfer experiments.
• Labeled splenocytes with carboxyfluorescein diacetate succinimidyl ester (CFSE) prior to intravenous injection.
• Employed the optical fractionator principle for microscopy-based quantification of CFSE-positive cells in thick organ sections (lung, spleen, liver, kidney, inguinal lymph node) at multiple time points (60 seconds to 1 week).

Main Results:

• Successfully obtained unbiased estimates of total transferred cell numbers across multiple organs.
• Quantified the distribution of CFSE-labeled cells from 60 seconds to 1 week post-transfer.
• Demonstrated the precision and simplicity of the optical fractionator method for cell enumeration.

Conclusions:

• The described method provides a direct and unbiased estimate of adoptively transferred cell numbers in various organs.
• This technique is well-suited for quantitative immunological studies requiring precise cell tracking.
• The optical fractionator principle offers a robust approach for assessing cell distribution and kinetics.