Motile chondrocytes from newborn calf: migration properties and synthesis of collagen II
Motile chondrocytes from newborn calf: migration properties and synthesis of collagen II
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DOI：
10.1016/s1063-4584(03)00087-6
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发表时间：
2003-08-01
影响因子：
7
通讯作者：
Morales, TI
中科院分区：
文献类型：
--
作者：
Chang, C;Lauffenburger, DA;Morales, TI
目的：确定分化的软骨细胞是否有运动能力。设计：将从六只动物中分离的小腿关节软骨细胞在转瓶中培养，并在第3天和第7天取出。进行Boyden室测定和延时视频显微镜以监测和量化细胞迁移。一种新的方法选择性收获和代谢标记的迁移细胞的基础上，细胞运动到下面的博伊登室膜。结果：在Boyden小室中，第3天软骨细胞在纤连蛋白包被的膜上的运动比在牛血清白蛋白包被的对照上的运动高3倍（分别为39+/-15vs12 +/-8cells/mm（2）（P=0.005））。胰岛素样生长因子-1（IGF-1，10 ng/ml）具有趋化性，可将运动性增加至87 +/- 16个细胞/mm（-）（与单独使用纤连蛋白的差异：P=0.0003）。对于第7天的细胞观察到类似的反应，但IGF-1活化并不明显（P=0.055）。由迁移细胞产生的胶原蛋白模式与标准II型胶原蛋白模式非常相似，没有任何胶原蛋白1产生的证据。在视频跟踪实验中，附着在纤连蛋白上的运动细胞表现出典型的板状伪足和丝状伪足，并且~ 30%的附着细胞是运动的（速度>1 μ m/h并且方向持久性> 1h）。典型的细胞路径长度为3G-50 pm，基本上大于一个完整的细胞长度displacement.Conclusion：一个人口分化良好的软骨细胞能够基质（COL II）的合成是能动的体外。这一最初的发现为研究运动细胞修复软骨的潜力开辟了新的途径。(C)2003年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: To determine whether differentiated chondrocytes are motile.Design: Calf articular chondrocytes isolated from six animals were cultured in spinner flasks and removed on days 3 and 7. Boyden chamber assays and time-lapse videornicroscopy were performed to monitor and quantify cell migration. A novel method for selectively harvesting and metabolically labeling the migrated cells was developed, based on cell movement to the underside of the Boyden chamber membranes. The 3 H-collagen synthesized by these cells was purified and analyzed by SDS-PAGE and autoradiography either before or after cyanogen bromide cleavage.Results: In Boyden chambers, locomotion of day 3 chondrocytes on fibronectin-coated membranes was similar to3-fold higher than on bovine serum albumin-coated controls (39+/-15 vs 12+/-8 cells/mm(2), respectively (P=0.005)). Insulin-like growth factor-1 (IGF-1, 10 ng/ml) was chemotactic, increasing motility to 87 +/- 16 cells/mm(-) (difference from fibronectin alone: P=0.0003). A similar response was observed for day 7 cells, but IGF-1 activation was not as pronounced (P=0.055). The collagen patterns produced by the migrated cells closely resembled those of standard collagen type II, without any evidence of collagen 1 production. In videotracking experiments, motile cells attached on fibronectin exhibited typical lamellipodia and filopodia, and -30% of attached cells were motile (speed >1 mum/h and directional persistence >1 h). Typical cell path lengths were 3G-50 pm, substantially greater than a full cell length displacement.Conclusion: A population of well-differentiated chondrocytes capable of matrix (COL II) synthesis are motile in vitro. This original finding opens new avenues to study the potential of motile cells for cartilage repair. (C) 2003 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.