CHONDROGENESIS OF LIMB BUD MESENCHYME INVITRO - STIMULATION BY CATIONS
CHONDROGENESIS OF LIMB BUD MESENCHYME INVITRO - STIMULATION BY CATIONS
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DOI：
10.1016/0012-1606(86)90252-6
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发表时间：
1986-06-01
影响因子：
2.7
通讯作者：
TUAN, RS
中科院分区：
文献类型：
--
作者：
ANTONIO, JDS;TUAN, RS
为了分析软骨形成中细胞-细胞相互作用的本质，我们在体外测试了影响这些相互作用的两种阳离子钙和聚赖氨酸（PL）对软骨形成的影响。用含钙（0.6 ~ 3.3 mM）或PL （1 ~ 10 μg/ml）培养基培养鸡肢芽间充质（Hamilton-Hamburger期23 ~ 24）高密度培养物。这两种阳离子都以剂量依赖的方式刺激软骨形成，并且在正常情况下无软骨形成、低细胞密度培养中也促进软骨形成。通过软骨结节数、[35 S]硫酸盐掺入和免疫组化检测II型胶原表达来检测软骨发生情况。其他二价阳离子（Cd, Co, Ni, Mg， Mn和Sr）没有模仿钙的作用。PL的效果依赖于它的M r（大于或等于14K）和电荷，并且由聚d-赖氨酸模拟，但不是由赖氨酸或PL或赖氨酸的其他类似物模拟（ϵ-amino己酸，溶菌酶，聚l-精氨酸和亚精胺）。钙和PL可能通过不同的机制起作用，因为它们的作用是加性的，并且需要它们在不同的培养天数存在：钙在第1天起作用，PL在第2天起作用。有人提出，钙可能在软骨形成的细胞聚集阶段发挥作用，而PL或一种天然存在的类似性质的多肽可能通过交联细胞表面或细胞外基质的特定阴离子组分来促进软骨形成。
To analyze the nature of cell-cell interactions in chondrogenesis, two cations that influence these interactions, calcium and poly-l-lysine (PL), were tested for their effects on chondrogenesis in vitro. High density cultures of chick limb bud mesenchyme (Hamilton-Hamburger stages 23 24), were exposed to culture media containing calcium (0.6-3.3 mM) or PL (1–10 μg/ml). Both cations stimulated chondrogenesis in a dose-dependent manner, and also promoted cartilage formation in normally non-chondrogenic, low cell density cultures. Chondrogenesis was assayed based on cartilage nodule number,[35 S] sulfate incorporation, and expression of type II collagen as detected by immunohistochemistry. The calcium effect was not mimicked by other divalent cations (Cd, Co, Ni, Mg, Mn, and Sr). The effect of PL was dependent on its M r (⩾ 14K) and charge, and was mimicked by poly-d-lysine but not by lysine or other analogs of PL or lysine (ϵ-amino caproic acid, lysozyme, poly-l-arginine, and spermidine). Calcium and PL probably act by different mechanisms since their effects were additive, and required their presence on different days of culture: calcium acted on Day 1, and PL on Day 2. It is proposed that calcium may play a role in the cell aggregation phase of chondrogenesis whereas PL, or a naturally occurring polypeptide of similar nature, may promote chondrogenesis by crosslinking specific anionic components of the cell surface or extracellular matrix.