Differentiation of rabbit amnion-derived stem cells induced into neural cells

Abstract

Objective To observe the differentiation effect of rabbit amnion-derived stem cells (ADSC) induced into neural cells.Methods ADSC of New Zealand female rabbits were isolated and cultured.Its mRNA level of Fibronectin,Nestin and Vimentin were detected by real-time quantitative polymerase chain reaction.The self-replication ability of ADSC was confirmed by monoclonal formation experiments.These ADSC were further induced into neural cells in vitro.Five days after induced differentiation,the expression of β-tubulin and glial fibrillary acidic protein (GFAP) were detected by immunofluorescent staining.Results ADSC were separated from amnion tissue gradually after 24 hours.There were polygonal cells gathered around the amnion tissue at 72 hours,and were distributed compactly around the amnion at 120 hours.The morphology of cleavage daughter cells was basically the same as parent cells.ADSC has the ability of self-replication.The Nestin,Vimentin,Fibronectin mRNA expressions in ADSC were 15.79,1.91,7.65 times those in spleen cells.The differences were statistically significant (Z=-5.243,-3.972,-2.524; P＜0.05).The β-tubulin expression was found in cytoplasm of most cells.The GFAP expression was found in cytoplasm in some cells.Conclusions ADSC has self-replication ability.It can be induced into neurons and neuroglial cells under the right conditions. Key words: Adult stem cells/cytology; Cell differentiation/immunology; Cell culture techniques/methods ; Amnion