Figure 3.

Mesenchymal stromal cells (MSCs) promote neural cell proliferation on extracellular matrix (ECM). Culturing on ECM supported proliferation of immature neurons. (A): Neural cell nuclei were counted in the presence and absence of MSCs. DAPI-stained paraformaldehyde-fixed cultures were assessed at a magnification of ×200. p values were calculated using the unpaired t test. (B): Illustration of morphological differences among DAPI-stained nuclei in a 7-day coculture allowing differential counting of nuclei: MSC nucleus (arrow), live neural cell nuclei (arrowheads), and dead (condensed) neural cell nuclei (asterisks). Magnification, ×400. (C): Cultures double-stained for BRDU and neuron-specific tubulin after a 7-hour incubation with BRDU on day 7. Magnification, ×200. The presence of BRDU⁺TUJ1⁺ cells indicated proliferation of neuronal precursors. Abbreviations: BRDU, 5-bromo-2′-deoxyuridine; DAPI, 4′,6-diamidino-2-phenylindole; M, mesenchymal stromal cells; N, neural cells; TUJ1, neuronal class III β-tubulin-specific antibody.