Identification of immune and non-immune cells in regenerating axolotl limbs by single-cell sequencing.

Identification of immune and non-immune cells in regenerating axolotl limbs by single-cell sequencing.
Exp Cell Res. 2020 Jun 17;:112149
Authors: Rodgers AK, Smith JJ, Voss SR
Abstract
Immune cells are known to be critical for successful limb regeneration in the axolotl (Ambystoma mexicanum), but many details regarding their identity, behavior, and function are yet to be resolved. We isolated peripheral leukocytes from the blood of adult axolotls and then created two samples for single-cell sequencing: 1) peripheral leukocytes (N = 7889) and 2) peripheral leukocytes with presumptive macrophages from the intraperitoneal cavity (N = 4998). Using k-means clustering, we identified 6 cell populations from each sample that presented gene expression patterns indicative of erythrocyte, thrombocyte, neutrophil, B-cell, T-cell, and myeloid cell populations. A seventh, presumptive macrophage cell population was identified uniquely from sample 2. We then isolated cells from amputated axolotl limbs at 1 and 6 days post-amputation (DPA) and performed single cell sequencing (N = 8272 and 9906 cells respectively) to identify immune and non-immune cell populations. Using k-means clustering, we identified 8 cell populations overall, with the majority of cells expressing erythrocyte-specific genes. Even though erythrocytes predominated, we used an unbiased approach to identify infiltrating neutrophil, macrophage, and lymphocyte popula...

Source: Experimental Cell Research - June 16, 2020 Category: Cytology Authors: Rodgers AK, Smith JJ, Voss SR Tags: Exp Cell Res Source Type: research