不同培养方式长期扩增神经干细胞的比较化学工程专业论文.docxVIP

不同培养方式长赣扩璜}}经干细胞的阮较Comparison 不同培养方式长赣扩璜}}经干细胞的阮较 Comparison ofDifferent Culture Mode for Long-term Expansion of Neural Stem Cells Abstract Neural stem cells附SCs)can be cultured in two modes of suspension and monolayer m vitro．In order to find the appropriate methods for la玛e_scale expansion ofNSCs，we compared the NSCs cultured in suspension with those culttwed in monolayer systematically．Firstly,the fombraln tissue was removed from the embryonic day 14∞14)mice,then the tissue was dissociated into single-cell suspension by AccutaseTM and mechanical trituration．The cells wg；e cultured in Ixah suspension and monolayer．The cells cultured in suspension and monolayer were identified协be NSCs by the assay of differentiation and immunofluorescence． Furthermore，the NSCs cultured in suspension and in monolayer were compared Oil viability, ability ofproliferation and heterogeneity by fluorescent dye，immunofluorescence and FACS on DIV21，DIV56 and DIVl 13 respectively．The results indicated that the NSCs cultured in both suspension and monolayer represented good viaU：iV in long—term culture．But they displayed distinct ability of proliferation in long—term culture．The NSCs cultured in monolayer preceded those cultured in suspension On the ability of proliferation 0n DIV21 and DIV56，but nO obvious difference on DIVt 13．The NSCs population cultured in suspension displayed more Nestin-positive cells than those in monolayer in the whole processes of culture．The NSCs population cultured in monolayer,however,displayed more t3tubulinlII-posifive cells than those in suspensioninthe Sameperiod．Additionally,wedeterminedthe suitabledlameterofneumspheresto be passed WSS about 200Ⅲn．The results of this study fnrther indicate that the cultured cells ale different In both the ability of proliferation and the heterogeneity．And the suspended culture modeexcelsthemonolayerculturemodeforlarge-scaleexpansionofNSCs。 Key Words：NSCs； suspension：monolayer 独创性说明作卷郑重声明：本硕士学位论文是我个人在导师指导F进行的研究工作 独创性说明 作卷郑重声明