Adult adipose-derived stem cells differentiation into a astrocyte cells morphology and ultrastructure in vitro / 中华行为医学与脑科学杂志

ABSTRACT

Objective To induce adult adult adipose-derived stem cells(ADSCs) in vitro to differentiate into neuronal-like cells,and to analyze the features of their cell morphology and ultrastructure. Methods Adipose stromal cells were obtained and amplified in vitro. Then make use of chemical induction to induce them. Observed ADSC and differentiation of cells in morphology and ultrastructure under inverted microscope and transmission electron microscopy. Immunocytochemistry to detection of Nestin, Neuron Specific Endolase( NSE) ,and glial fibrillary acidic protein(GFAP) expression in cells. Used Real-time PCR to detection of Nestin,GFAP gene mRNA expression before and after induction in ADSC. To observe the morphology and ultrastructure of the cells prior to and after induction under microscope and electron microscope. Results The morphology of ADSC was similar to fibroblasts ,and could be amplified stability within 10 passages in vitro. Some of the cells induced display a typical astrocyte-like cells in ultrastructure. Followed neuronal induction,astrocyte-like cells began to stain brightly for CFAP, Nestin. GFAP stained in both the cytoplasm and nuclei of astrocyte-like cells, but Nestin only stained in the cytoplasm. The peak positive expression rate within 14d following neuronal induction. The rate of positive expression cells was( 14.4 ± 3. 6) % for Nestin, (87. 3 ± 5. 3 ) % for GFAP. Then two kinds of protein expression remained the similar rate. The average relative concentration of GFAP and Nestin gene mRNA have significant statistical difference between ADSC and differented cells analyzed by Real-time PCR (P<0.05).The peak concentration of GFAP was within 20 d after induction,and GFAP was within 14 d after induction. Conclusion In the cytoplasm of adult adipose-derived cells possess Nestin genetic material,which is the marker of neural stem cell. The differential astrocyte-like cells have the typical morphology, ultrastructure and GFAP phenotype of mature astrocytes.