Osteogenic characteristics of Naringin-induced rabbit bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2013.14.020

Abstract

Abstract:

BACKGROUND: Drynaria can promote bone growth and has made good clinical curative effect. Can the main ingredients of Naringin induce bone marrow measenchymal stem cells to differentiation into osteoblasts?
OBJECTIVE: Through the use of Naringin to induce osteogenic differentiation of rabbit bone marrow mesenchymal stem cells, to observe the osteogenic ability of Naringin-induced rabbit bone marrow mesenchymal stem cells.
METHODS: Adherence screening method was used to separate and culture the rabbit bone marrow mesenchymal stem cells. The well-grown third generation of bone marrow measenchymal stem cells were induced with 50 ug/L Naringin and classic osteogenic inducer respectively to differentiate into the osteoblasts, and the osteogenesis identification was performed.
RESULTS AND CONCLUTION: Both the classic osteogenic inducer and Naringin induced liquid could induce the bone marrow mesenchymal stem cells to differentiation into osteoblasts with increasing matrix secretion and calcium nodules formation. The enzyme-linked immunosorbent assay showed that there was no significant difference in absorbance value of the cells between the Naringin induction group and the classic osteogenic inducer group. There was no significant difference in absorbance value of the cells induced with Naringin induced liquid and L-DMEN culture medium (P > 0.05). The results indicate that Naringin can induce the bone marrow mesenchymal stem cells to differentiate into osteoblasts without obvious toxic effect.

Key words: stem cells, stem cell culture and differentiation, Naringin, bone marrow mesenchymal stem cells, osteoblasts, cell differentiation, osteogenic induction, cell culture, alkaline phosphatase, collagen type Ⅰ, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Yang Yuan, Li Xiao-feng, Luo Dao-ming, Wen Chao-hai. Osteogenic characteristics of Naringin-induced rabbit bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(14): 2603-2608.

Figures/Tables 4

2.1 兔骨髓间充质干细胞原代培养结果刚接种的细胞镜下呈圆形，大小较均一，24 h后见部分贴壁的细胞呈圆形、椭圆形、立方形、对角形等不规则性[8-9]，体积较悬浮的圆形细胞大，大小较均一，其核浆分界清楚，折光性强，绝大多数为单核、圆形，位于细胞中央。48 h后全量更换培养液，细胞逐渐铺开，呈成纤维细胞样克隆集落贴壁生长，呈放射状，直径大小不等。细胞轮廓呈长梭形，少量细胞呈星形，胞浆丰富，常有一两个细胞核，居中。随着培养时间的延长，这些贴壁细胞形成集落，集落大小不一，培养液中还有小圆型的细胞极形。细胞增殖迅速，集落之间相互靠进，最终细胞融合成单层，细胞梭形突起变长，排列有明显的方向性，呈漩涡状。 2.2 兔骨髓间充质干细胞传代细胞培养结果传代细胞接种后6-8 h贴壁，而后逐渐伸展为梭形、多角形。传代后的细胞生长迅速，两三天可基本铺满培养瓶底。原代培养发现局部区域少许小而圆的细胞，经传代后消失。传代后细胞形态上无明显改变，随着传代次数的增加细胞开始老化，表现为细胞碎片的增多，细胞表现出明显的衰老现象，细胞增殖速度明显减慢，细胞形态变为扁平，细胞活力低下，遮光性差，增殖速度明显减慢直至停止[10]。 2.3 兔骨髓间充质干细胞生长曲线以时间(天)为横坐标，每天取6孔细胞吸光度值的平均值为纵坐标绘制细胞生长曲线图。传代后细胞生长曲线大致相同，细胞接种后一两天内数量无明显变化，第2天比第1天的细胞数量甚至还稍有减少。第3天起，进入对数增殖期，细胞大量增加，至第7天细胞数量达到高峰，以后细胞增长速度减慢，见图1。细胞倍增时间约为45 h。"

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