Effect Of Self-assembling Nanofiber Gel Containing IKVAV On The Proliferation Of Neural Stem Cells And Differentiation Into Neurons In Vitro

Objective: To observe the effects of self-assembled IKVAV nanofiber gel onproliferation and differentiation of bone marrow derived NSCs in vitro as well as itscytotoxicity.Methods: A stem cell group, a gel group, and a gel-stem cell combininggroup were set for comparison. The cultured cells were identified throughimmunocytochemistry with anti-Nestin antibody using the SP method; cell proliferationrate was measured at1,3,7,14days after establishment of each experimental group bythe CCK-8method; MTT assay was used for detection of cytotoxicity; and flow cytometrywas used to calculate the differentiation rate of NSCs into neurons and to analyze cellcycle.Results: For co-culture group, cell aggregation was seen in1day; neurospheresformed and gradually grew in3days and reached a diameter of100um on day7. Theneurospheres appeared plump, and mulberry-like particles formed at their edge. Nestinstaining was positive. CCK-8assay results showed that: cell proliferation rate ofco-cultured group was significantly higher than that of gel group and stem cell group(P<0.05); cell proliferation rate of both the co-culture group and the stem cell groupreached a peak at day7(0.58±0.02,0.4467±0.2517respectively) and no cell proliferationwas detected for the gel group. MTT assay detected no cytotoxicity of the gel (P=0.82).Flow cytometry results showed that differentiation proportion of NSC into neurons wassignificantly higher in the co-culture group than the other two groups at each time points(P<0.05). Differentiation proportion at1d,3d,5d,7d,10d,14d were2.1%,3.0%,4.6%,7.1%,5.8%,2.7%for the co-culture group and1.9%,2.3%,3.5%,4.7%,3.1%,1.6%forthe stem cell group; no neurons was detected in gel group.Conclusion: IKVAV nanofibergel had good biological activity, can increase the proportion of NSCs into neurons.