Stem Cell Research Center, Anatomy Department, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Glycoconj J. 2011 Feb;28(2):89-98. doi: 10.1007/s10719-011-9326-5. Epub 2011 Mar 1.
The role of cell surface glycoproteins in cell behavior can be characterized by their interactions with plant lectins. This study was designed to identify the effects of lectins on chondrogenesis and osteogenesis in limb bud mesenchymal cells in vitro. Limb bud mesenchymal cells from mouse embryos were cultured in high-density micromass culture. Wheat germ agglutinin (WGA), concanavalin A (ConA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA) and Ricinus communis agglutinin (RCA) were added separately to the culture media. Cells were cultured for 5 or 9 days, and cell viability was assayed by neutral red on day 5. The micromasses were stained with alcian blue, alizarin red S and Von Kossa stains, and alkaline phosphatase assays were also done. Dolichos biflorus agglutinin induced an increase in chondrogenesis, calcium precipitation and proteoglycan production. ConA and PNA did not affect chondrocyte differentiation but induced chondrocytes to produce more proteoglycan. Wheat germ agglutinin reduced chondrification and ossification but induced mesenchymal cells to store lipid droplets. Ricinus communis agglutinin 1 was toxic and significantly reduced cell survival. In conclusion, DBA was the most effective inducer of ossification and chondrification. Wheat germ agglutinin induced adipogenesis instead. These assays showed that lectins play important roles in limb bud development.
细胞表面糖蛋白与植物凝集素的相互作用可以用来描述其在细胞行为中的作用。本研究旨在确定凝集素对体外肢芽间充质细胞软骨生成和骨生成的影响。从小鼠胚胎肢芽中分离出间充质细胞,在高密度微团培养中进行培养。分别向培养基中添加麦胚凝集素(WGA)、伴刀豆球蛋白 A(ConA)、花生凝集素(PNA)、双花扁豆凝集素(DBA)和蓖麻凝集素(RCA)。培养 5 或 9 天,第 5 天通过中性红测定细胞活力。用阿利新蓝、茜素红 S 和Von Kossa 染色对微团进行染色,并进行碱性磷酸酶测定。双花扁豆凝集素诱导软骨生成、钙沉淀和蛋白聚糖产生增加。ConA 和 PNA 不影响软骨细胞分化,但诱导软骨细胞产生更多的蛋白聚糖。麦胚凝集素减少软骨形成和骨化,但诱导间充质细胞储存脂滴。蓖麻凝集素 1 具有毒性,显著降低细胞存活率。总之,DBA 是最有效的成骨和软骨诱导剂。麦胚凝集素诱导脂肪生成。这些实验表明,凝集素在肢芽发育中起重要作用。