Experimental study on differentiation of different passages of human bone marrow mesenchymal stem cells into neurons in vitro
- VernacularTitle:不同代数成人骨髓间充质干细胞体外转化为神经细胞的实验
- Author:
Ying CHANG
;
Xin QI
;
Hong YANG
;
Ping XU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2005;9(46):130-132
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Bone marrow mesenchymal stem cell (MSC) is a kind of stem cell with potential of self-repair and multi-differentiation. It may differentiate into neuron, adipose cell and osteoblasts.OBJECTIVE: To observe the transforming efficacy of human bone marrow mesencymal stem cells (hMSCs) into neurons in vitro in different generations so as to provide reliable experimental data for the clinical application of MSCs.DESIGN: Single sample was designed.SETTING: Department of Neurology, Sino-Japan Friendship Hospital,Jilin UniversityPARTICIPANTS: Marrow tissue was collected from 9 cases of spinal fusion in Department of Orthopedics of First Hospital affiliated to Jilin University. Of 9 cases, all of them were in known of the experiment.METHODS: The experiment was performed in Sino-Japan Hospital affiliated to Jilin University from September 2002 to February 2003. The primary and generative culture of hMSCs was given. Experimental and the control groups were divided.-mercaptoethanol was taken as inducer. hMSCs of the 2rd, 4th, 6th and 8th generations were selected for induction in vitro for 6 h. Cytochemistry staining and immunohistochemistry staining were used to assay the expressions of neuronal and astrocytic marked proteins.MAIN OUTCOME MEASURES: ① Growth curve analysis on genera tive culture of hMSCs. ② Nissel staining. ③ Immunohistochemical staining.RESULTS: ① Common characters of generative culture: The latent phase of generative culture was 12-24 hours, exponential phase was 7-10 days and 11-13 days later, cell culture entered the platform phase. ② After induction of the 2nd, 4th and 6th generated hMSCs, deep blue granular Nissl body presented in cytoplasm. In 6 hours on the 8th induction, there was no obvious deep blue Nissl structure presented in cytoplasm.③ Except GFAP, NSE and NF-M were expressed in hMSCs of different generations after induction for 6 hours. There was no significant difference in positive rates of the 2nd, 4th and 6th generations (P ＞ 0.05), but the significant difference presented in comparison between the 8th generation and the 2nd, 4thand 6th generations (P ＜ 0.05). CONCLUSION:-mercaptoethanol can induce hMSCs differentiating into neuronal cells in vitro. The positive rates of the 2nd, 4th and 6th generationsare higher remarkably than the 8th generation.
