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Huperzine A protects neural stem cells against A?-induced apoptosis in a neural stem cells and microglia co-culture system.

ABSTRACT: This study aims to explore whether Huperzine A (HupA) could protect neural stem cells against amyloid beta-peptide A? induced apoptosis in a neural stem cells (NSCs) and microglia co-culture system.Rat NSCs and microglial cells were isolated, cultured and identified with immunofluorescence Assays (IFA). Co-culture systems of NSCs and microglial cells were employed using Transwell Permeable Supports. The effects of A?1-42 on NSCs were studied in 4 groups using co-culture systems: NSCs, A?+NSCs, co-culture and A?+co-culture groups. Bromodeoxyuridine (BrdU) incorporation and flow cytometry were utilized to assess the differences of proliferation, differentiation and apoptosis of NSCs between the groups. LQ test was performed to assess the amounts of IL-6, TNF-? and MIP-? secreted, and flow cytometry and Western blotting were used to assess apoptosis of NSCs and the expressions of Bcl-2 and Bax in each group.IFA results showed that isolated rat NSCs were nestin-positive and microglial cells were CD11b/c-positive. Among all the groups, the A?+co-culture group has the lowest BrdU expression level, the lowest MAP2-positive, ChAT-positive cell counts and the highest NSC apoptosis rate. Smaller amounts of IL-6, TNF-? and MIP-? were being secreted by microglial cells in the HupA+A?+co-culture group compared with those in the A?+ co-culture group. Also the Bcl-2: Bax ratio was much higher in the HupA+A?+co-culture group than in the A?+co-culture group.HupA inhibits cell apoptosis through restraining microglia's inflammatory response induced by A?1-42.

SUBMITTER: Zhu N

PROVIDER: S-EPMC4525852 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Huperzine A protects neural stem cells against Aβ-induced apoptosis in a neural stem cells and microglia co-culture system.

Zhu Ning N Lin Jizong J Wang Kewan K Wei Meidan M Chen Qingzhuang Q Wang Yong Y

International journal of clinical and experimental pathology 20150601 6

<h4>Objectives</h4>This study aims to explore whether Huperzine A (HupA) could protect neural stem cells against amyloid beta-peptide Aβ induced apoptosis in a neural stem cells (NSCs) and microglia co-culture system.<h4>Methods</h4>Rat NSCs and microglial cells were isolated, cultured and identified with immunofluorescence Assays (IFA). Co-culture systems of NSCs and microglial cells were employed using Transwell Permeable Supports. The effects of Aβ1-42 on NSCs were studied in 4 groups using c ...[more]

PMID: 26261518

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Project description:Embryonic stem cells (ESCs), due to their intrinsic capability to generate somatic cells of all three germ layers, are potential sources of neural cells for cell replacement therapies. However, the empirical differentiation protocols and the lack of mechanistic understanding of the neural differentiation of ESCs have limited the utility of ESCs as a developmental model or as a cell source for neural cell populations for replacement therapies. Co-culturing ESCs with stromal cells is one of the extensively used methods to induce neural differentiation. Despite several studies to identify neural inducing factors in stromal cell induced neural differentiation, the self-regulatory effects of ESCs in the neural differentiation process remain unexplored. For the first time, we elucidate the self-regulatory role of mESCs in their neural cell differentiation by supplementing conditioned media from differentiating mESCs to mESC-PA6 co-cultures and quantitatively evaluating the change in neural differentiation. Moreover, we use statistical tools to analyze the expression of various growth and trophic factors and distinguish the factors produced primarily by PA6 cells versus mESCs in co-culture. We observe that addition of the medium containing mESC-secreted factors to a single mESC colony co-cultured with PA6 cells significantly enhances the neural differentiation of mESCs compares to the medium extracted from the stromal cells only. Hierarchical clustering of gene expression data from PA6 and co-cultured mESCs segregates two groups of factors that are produced by the stromal cells and differentiating mESCs. Identifying the major soluble factors that drive and regulate the neural differentiation process in the mESC-PA6 co-culture niche will help understand molecular mechanisms of neural development. Moreover, it can be a major step toward developing novel protocols to differentiate stem cells with mESC derived factor supplementation without using feeder cells and with greater efficiency compared to existing approaches.

Dataset Information

Huperzine A protects neural stem cells against A?-induced apoptosis in a neural stem cells and microglia co-culture system.

Publications

Huperzine A protects neural stem cells against Aβ-induced apoptosis in a neural stem cells and microglia co-culture system.

Similar Datasets

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