Establish And Evaluation Of Human Brain Slice Culture To Expand Neuroscience Research

Objective: To establish and optimize an appropriate culture method ofbrain tissue in vitro, and to prepare reliable brain slices for electrophysiologicalexperiments.Methods:(1) Establish a stable brain slice culture method of young Wistarrats in vitro: To Optimization the culture techniques of rodent brain slice, weimproved the compositions of artificial cerebrospinal fluid (ACSF), and usedACSF with different ingredients in the process of slicing and culture withsubmerged slice culture-high flow rate technique. Dynamic changes of localfield potentials (LFP) were monitored for48h and histological and pathologicalchanges of brain slice were observed using HE staining, triphenyltetrazoliumchloride (TTC) staining and Nissl staining.(2) Human brain slice culturemethod： Based on the optimized rodent brain slice culture method, thenon-pathological cortex, removed from the excision surgery for human deeperbrain tumor was sliced, transferred and cultured rapidly. The LFP and the abovehistological and pathological indicators were also observed in human brainslices to evaluate the culture method.Results:(1) The LFP electrophysiological signals of young rats brain sliceswere sustainable at least48hours, human brain slices as well. But following thetime of the elongation the amplitude of LFP signal was declined.(2) Countedthe mortality rate of slice neurons by HE staining under light microscope, bothhuman and rat brain slice neurons could maintain more than50%survival ratefor24h, rats neuron survival rate was slightly higher than that of human brainslices.(3) IOD values obtained by TTC staining at each time point present a decreasing trend, the red stained region was still visible at48h, but both of thecomparisons were without statistical significance (P>0.05).(4) IOD values byNissl staining present a decreasing trend, both rat and human brain slices beganto decrease at the24h time point, dropped to a lower level at48h.Conclusions: As similar with rodent brain slice in cultured survival time,human’s, when sliced, could keep more than50%neurons survival more than24h, which could satisfy the neural electrophysiology research requirement ofliving cells. Using different ACSF solutions, and submerged slice culture-highflow rate technique, could meet that it need about1h transportation periodbefore the non-pathological cortex tissue removed from surgery processing.