# Identification and Ultrastructural Peculiarities of Nestin-Carrying Progenitor Cells in Kidney

## TL;DR

This study identifies and examines the unique structure of nestin-positive progenitor cells in kidney cultures using electron microscopy.

## Contribution

The study reveals ultrastructural differences between nestin-positive and nestin-negative kidney cells using GFP and electron microscopy.

## Key findings

- Nestin-positive cells showed high ribosome content and protein-synthesizing activity.
- Nestin-positive cells had vesicle-containing protrusions and multiple nuclei with high lysosome content.
- Tight interactions and structural differences were observed between nestin-positive and -negative cells.

## Abstract

In this study, in a culture of renal epithelial cells, we identified those expressing nestin, a cytoskeletal protein associated with stem/progenitor/activated/proliferating cell states. A mouse expressing GFP under the nestin promoter was used, followed by cell isolation and culture. It is hypothesized that this can be used to assess the stem/progenitor/activated/proliferating cell level in a mixed kidney cell culture. Both nestin-positive and nestin-negative cells were demonstrated to be present in the culture. After visualization, cells were attached to a glass slide with a grid, fixed, and prepared for electron microscopy analysis, with each cell visually identified by light microscopy being analyzed. Electron microscopy revealed tight interactions between nestin-positive and nestin-negative cells. Significant differences in the ultrastructure of nestin-positive and nestin-negative cells were observed. Nestin-positive cells were distinguished by a high ribosome content, indicating high protein-synthesizing activity. In the nestin–GFP-high (sorted) population examined by electron microscopy, vesicle-containing protrusions were frequently observed. These cells could contain multiple nuclei of varying sizes and had a high content of lysosomes. No significant differences in mitochondrial ultrastructure were observed in nestin-positive and -negative cells, although functional characteristics evaluated by the membrane potential probe differed.