Induction Of Human Bone Marrow Mesenchymal Stem Cells Differentiate Into Photoreceptor Cells In Vitro

OBJECTIVE: To explore the microenvironment which BMSCs differentiated into photoreceptor cells under nutrilits induction medium in vitro in order to open up a new way for clinical to cure retinal degenerative disease.METHODS: Bone marrow obtained from five healthy adult donors following informed consent. High-purity BMSCs were obtained using Lymphocyte Separation Medium by density gradient centrifugation. Flow cytometry was employed to detect BMSCs surface antigens. In the first stage the third passage cells were induced to neural precursor cells with medium including bFGF, EGF and BDNF. The expression of the Nestin and the MAP-2 were detected by immunocytochemistry. PEDF and Taurine instead of bFGF, EGF and BDNF in medium cultured the induced BMSCs furthermore 2 weeks when the positive expression rate of Nestin reached the highest level. Rhodopsin was researched during this second stage by immunocytochemistry and RT-PCR.RESULTS: Nestin could be detected since the third day of inducing differentiation, and reached its peak on the 12th day,(90.9±2.6)%, then the positive rate dropped down t to(75.5±3.7)% on the 14th day. MAP-2 also could be detected since the sixth day after inducing. Rhodopsin was detected during the farther inducing, and the positive rate was(20.7±3.8)% in the second week.CONCLUSION: In the first stage of induction BMSCs could differentiate into the nerve precursor cells which could express the Nestin and the MAP-2 under the condition of bFGF, EGF, BDNF,and in the second stage of induction under the condition of PEDF and Taurine the cells could express Rhodopsin which is the marker of photoreceptor cells. This research provides new ideas for cure retinal degenerative diseases.