Human Brain Organoids Derived Mesenchymal Stem Cells Promotes Functional Recovery In Alzheimer’s Disease Mouse

Objective The study aim to investigate the effect of human brain organoids derived mesenchymal stem cells and its treatment effect on Alzheimer’s disease(AD)mice.Method 1)The acquisition of neural derived mesenchymal stem cells(N-MSCs):A stable cell line was isolated from floating cultured brain organoids and was named N-MSCs.N-MSCs were characterized and confirmed by morphological observation,flow cytometry and three-way differentiation ability test.2)Compared gene expression of N-MSCs with other MSCs,including dental pulp MSC(P-MSCs),umbilical cord MSCs(UB-MSCs),bone marrow MSCs(BM-MSCs),iPSC derived directly MSCs(iPSCMSCs)and neural crest cell.3)28-week-old AD male mice were randomly allocated into three groups for cell injection therapy(PBS group,UB-MSCs group,and N-MSCs group).Each treatment was injected 0.2mL cell suspension(1×106 cells/mL)via tail vein for 8 weeks.Behavioral tests include Open field test,New object recognition test,Fear conditional test,and Barnes maze test(n=9).4)Histological evidence of N-MSCs improved AD mice symptom was evaluated by newborn neurons,dendritic spines of neurons,neural stem progenitors in the hippocampus,amyloid plaques and microglia cells.5)Omics Technology was used to explore mechanism.Results 1)N-MSC was similar to UB-MSCs and identified as a kind of MSCs.2)Compared with MSCs from other sources,the differential genes of N-MSCs were highly correlated with neuronutrition,inflammatory response and nervous system development.3)In the Open field test,AD mice in the N-MSCs group showed more exploration behavior.In the New object recognition test,compared with the PBS group,the UB-MSCs group and the N-MSCs group scored higher in exploring new objects,but only N-MSCs group achieved the passing score.In the Fear conditional experiment,mice in the N-MSCs group showed stronger learning ability and more Freezing behavior.In the Barnes maze test,N-MSCs group mice showed the shortest escape latency in Dayl Train l and Day 12 test,which showed stronger learning ability and better long-term memory.4)The section of AD mice brain hippocampal region showed the longest and the most complexity of newborn neurons in the N-MSCs group.More neural stem cells were found in N-MSC group DG region.The number of mature mushroom dendrites spine and the density of dendritic spines at the end of neurons were the highest in N-MSC group.The differences were statistically significant.The number of amyloid plaques(area>100 μm2)in the DG and CA1 region of N-MSC group mice was significantly lower than that in the PBS group(P<0.05).The morphologies of microglia in N-MSCs group tended to be inactive and shorter branching length than PBS group and UB-MSCs group.5)The omics analysis showed N-MSCs up-regulated inflammation regulation and neurotrophin signaling pathway.Conclusion N-MSCs can improve the symptoms of AD mice.