Conditioned medium of human menstrual blood-derived endometrial stem cells protects against cell inflammation and apoptosis of Npc1KO N2a cells

AbstractNiemann-Pick disease type C1 (NPC1) is a hereditary neurodegenerative disorder caused by a mutation in theNPC1 gene. This gene encodes a transmembrane protein found in lysosomes. This disease characterized by hepatosplenomegaly, neurological impairments and premature death. Recent preclinical studies have shown promising results in using mesenchymal stem cells (MSCs) to alleviate the symptoms of NPC1. One type of MSCs, known as human menstrual blood-derived endometrial stem cells (MenSCs), has attracted attention due to its accessibility, abundant supply, and strong proliferation and regeneration capabilities. However, it remains uncertain whether the conditioned medium of MenSCs (MenSCs-CM) can effectively relieve the symptoms of NPC1. To investigate this further, we employed the CRISPR-Cas9 technique to successfully create aNpc1 gene knockout N2a cell line (Npc1KO N2a). Sanger sequencing confirmed the occurrence ofNpc1 gene mutation in these cells, while western blotting revealed a lack of NPC1 protein expression. Filipin staining provided visual evidence of unesterified cholesterol accumulation inNpc1KO N2a cells. Moreover,Npc1KO N2a cells exhibited significantly decreased viability, increased inflammation, and heightened cell apoptosis. Notably, our study demonstrated that the viability ofNpc1KO N2a cells was most significantly improved after being cultured by 36  h-collected MenSCs-CM for 0.5 days. Additionally, MenSCs-CM exhibited the ability to effectively red...
Source: Metabolic Brain Disease - Category: Neurology Source Type: research