JIANG Xiaorui 1 , ZHANG Xinxin 1 , YANG Keyue 1 , JIANG Shan 1 , JIN Dan 1 , WANG Dan 1 , PEI Guoxian 2
  • 1Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou Guangdong, 510515, P.R.China;;
  • 2Institute of Orthopedics of Chinese PLA, Xijing Hospital, Fourth Military Medical University.;
Bone marrow mesenchymal stem cells; Quantum dot; Label ing in vitro; Rat
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Objective To explore the cytotoxicity, labeled time, marking rate, and effect on adhesion of quantum dot 655 (QD655) labeled rat bone marrow mesenchymal stem cells (BMSCs) in vitro, and to confirm its feasibil ity for stem cell label ing and tracer means for rat. Methods BMSCs were collected from the femur and tibia bone marrow cavity of a 2-week-old SD rat, cultured and identified. The 3rd passage of BMSCs were incubated with QD655 as the experimental group
according to the recommended concentration of the markers. The cells were not labeled by QD655 as control group. The
cell survival rate after QD655 label ing was detected by trypan-blue exclusion. The effect of QD655 on cell prol iferation was observed by MTT. The osteogenic differentiation potential was identified by Al izarin red staining, alkal ine phosphatase (ALP) staining, and real-time fluorogenic quantitative PCR. At immediately, 1, 2, 4, and 6 weeks, fluorescent microscopy was used to observe the labeled rate and scanning electron microscope was used to observe the cell adhesion to scaffold (bioglass/collagen composite). Results The cell survival rates were more than 90% in both experimental group and control group, showing no significant difference (P  gt; 0.05). There was no significant difference in the cell prol iferation between 2 groups (P  gt; 0.05). Al izarin red staining and ALP staining showed positive results. Real-time fluorogenic quantitative PCR result showed that the mRNA expression levels of osteopontin, osteocalcin, collagen type I, ALP, and BMP-2 in the experimental group was significantly higher than those in the control group. The labeled rates were 96.50% ± 1.59%, 93.30% ± 1.51%, 72.40% ± 2.90%, 40.10% ± 3.60%, and 10.00% ± 1.70% immediately, 1, 2, 4, and 6 weeks after label ing, respectively. The labeled rate in the control group was 0. Scanning electron microscope showed a good distribution of fusiform or polygonal cells in the pores of scaffold. Conclusion QD655 can be used as a label ing marker for BMSCs. Rat BMSCs labeled with QD655 is of high efficiency and safety.

Citation： JIANG Xiaorui,ZHANG Xinxin,YANG Keyue,JIANG Shan,JIN Dan,WANG Dan,PEI Guoxian. IN VITRO QUANTUM DOT-LABELED RAT BONE MARROW MESENCHYMAL STEM CELLS. Chinese Journal of Reparative and Reconstructive Surgery, 2010, 24(6): 744-748. doi: Copy