Effect of Acorus tatarinowii on the osteogenic differentiation of umbilical cord blood stem cells

doi:10.3969/j.issn.2095-4344.2016.41.008

Abstract

Abstract:

BACKGROUND: Studies have demonstrated that Acorus tatarinowii and its active ingredients can promote adult neurogenesis, exerting an active role in anti-aging and neurodegenerative disease treatment.
OBJECTIVE: To explore the effects of Acorus tatarinowii extracts on the proliferation and osteogenic differentiation of umbilical cord blood stem cells, thereby providing a new idea for promoting the osteogenic differentiation of stem cells by Chinese medicines.
METHODS: Acorus tatarinowii extracts were obtained via solvent extraction method and flow cytometry sorting technology was used to select the stem cells isolated from human umbilical cord. Then, the umbilical cord blood stem cell proliferation was observed by electron microscope, and the effect of Acorus tatarinowii on the proliferation of umbilical cord blood stem cells was observed by cell counting kit-8. Meanwhile, the impact of Acorus tatarinowii on osteocalcin and bone morphogenetic protein-2 contents in the supernatant of umbilical cord blood stem cells were detected by ELISA; alkaline phosphatas expression was detected using alkaline phosphatase staining kit.
RESULTS AND CONCLUSION: The separation purity of the stem cells from umbilical cord mononuclear cells was (89.66±3.47)%. After low, moderate and high concentrations of cord blood stem cells co-cultured with Acorus tatarinowii extractions for 24, 48 and 72 hours, the stem cell proliferation rate was significantly higher compared with the control group (P < 0.05), and additionally, the proliferation rate of moderate concentration group was significantly higher than that in the low and high concentration groups (P < 0.05). The contents of osteocalcin and bone morphogenetic protein-2 in the stem cell supernatants were significantly higher than those in the control group after co-cultured with Acorus tatarinowii extractions for 5, 10 and 15 days, and which the highest in the high concentration group (P < 0.05). The expression of alkaline phosphatase was significantly higher than that in the control group after umbilical cord blood stem cells co-cultured with Acorus tatarinowii extractions for 10 days, and moreover, the expression of alkaline phosphatase in the moderate concentration and high concentration groups were significantly higher than that in the low concentration group (P < 0.05). In conclusion, Acorus tatarinowii can promote the proliferation and osteogenic differentiation of umbilical cord blood stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Drugs, Chinese Herbal, Bone Morphogenetic Proteins

CLC Number:

R394.2

Chu Jing, Wang Li-qin, Zhang Han, Zeng Juan . Effect of Acorus tatarinowii on the osteogenic differentiation of umbilical cord blood stem cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(41): 6131-6137.

Figures/Tables 3

2.1 脐血干细胞的提取分离及培养从脐血中分离得到的单核细胞平均数量为(1.68±0.32)×1010 L-1，经磁珠分选得到的干细胞数量为(1.78±0.14)×108 L-1，经流式细胞技术鉴定发现所得脐血干细胞纯度达到(89.66±3.47)%(见图1)。对分选出的脐血干细胞进行培养，刚分离的细胞呈圆形，分离7 d后，贴壁生长，呈梭形，分离14 d的细胞长满培养瓶，见图2。 2.2 石菖蒲对脐血干细胞增殖的影响 CCK8结果显示，加入石菖蒲浸提液低剂量、中剂量、高剂量培养脐血干细胞24，48，72 h后，脐血干细胞的增殖率明显高于空白对照(P < 0.05)，其中，中剂量组的增殖率明显高于低剂量、高剂量组(P < 0.05)，低剂量与高剂量组间的增殖率差异无显著性意义(P > 0.05)。见表1。 2.3 石菖蒲对脐血干细胞培养上清液中骨钙素含量的影响由ELISA实验结果可知，加入石菖蒲浸提液低剂量、中剂量、高剂量培养脐血干细胞5，10，15 d后，上清液中骨钙素的含量明显高于空白对照组(P < 0.05)，其中，中剂量组的骨钙素含量高于低剂量、高剂量组 (P < 0.05)。见表2。 2.4 石菖蒲对脐血干细胞培养上清液中骨形态发生蛋白2含量的影响由ELISA实验结果可知，加入石菖蒲浸提液低剂量、中剂量、高剂量培养脐血干细胞5，10，15 d后，上清液中骨形态发生蛋白2的含量明显高于空白对照组(P < 0.05)，其中，中剂量组的骨形态发生蛋白2含量高于低剂量、高剂量组(P < 0.05)。见表3。 2.5 碱性磷酸酶染色检测石菖蒲对脐血干细胞中碱性磷酸酶分布表达的影响碱性磷酸酶染色结果显示，加入石菖蒲浸提液培养干细胞10 d后，低剂量、中剂量、高剂量组中碱性磷酸酶的分布表达明显高于空白对照，中剂量组和高剂量组的表达明显高于低剂量组。见图3。"

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