Experimental Research On The Spin's Regeneration Using BMSC And FK506 After SCI In Vitro

Experimental Research on the Spin's Regeneration Using BMSC and FK506 afterSCI in vitro.[Objective] To investigate the effect of bone mesenchymal stem cell (BMSC) andtacrolimus (FK506) on spinal cord injury (SCI).[Method] Primary culture of rat BMSC were collected from the femurs andtibias of one month old SD rat. The bone marrow was washed from marrowcavity with L-DMEM complete medium. After filtrated, the marrowsuspension were collected and placed in a 25cm² flask. The culture weremaintained at 37℃, 5% CO₂ and saturated humidity. The non-adherent cellswere removed by replacing the medium after 72 hours of culture andthereafter to replace the medium every 72 hour. When the cells had grownto confluence(about 14 days), they were passaged by 0.25% trysin andreplated after dilution to 1:2 or 1:3. The passaged rBMSC attached toplastic flask with in 12 hours and reached confluence within 7 to 10 daysfor transplantation. The ring clamp SCI model of rat was constructed, 30 ratswere divided randomly into three groups, group A was single SCI group, group B wasSCI group treated with BMSC injected to the area of SCI, group C was SCI grouptreated just like group B but with FK506 which injected through abdominal part. In 1,2, 4,6,8 weeks post-injury, all animals were evaluated of the hind limb behaviorwith BBB score and inclined plane test. Animals showed vary degree of functionalrecovery. The immunohistochemistry and pathology were checked after 8 weeks.[Result] All the rats hind limb function were all recovery obviously, the hind limbs ofgroup B and C begin to move and they start to urinate gradually after 2 weeks,especially after 4 weeks their hind limbs and tails could move actively. The outcomeof C group surpass the B group's, the highest BBB scores were 14. There wassignificant difference between B and C group(P＜0.05). The A group's outcome weresimilar to B and C, but it's degree was smaller than the others'. There was significantdifference between B,C and A group after 6 weeks also (P＜0.05).[Conclusion] The functions of hind limbs of rats with complete SCI can recoverobviously, the BMSC which be planted survival well in the area of injury cord, andcan transfer to the surroundings. The BMSC possess promotion effects on injuredspinal cord in rats, with the affiliation of FK506 claim synergism effects. FK 506 isone of the advantage respects which the recovery of injured cord need.