HEPATOCYTE GROWTH-FACTOR IMMOBILIZED ONTO CULTURE SUBSTRATES THROUGH HEPARIN AND MATRIGEL ENHANCES DNA-SYNTHESIS IN PRIMARY RAT HEPATOCYTES
HEPATOCYTE GROWTH-FACTOR IMMOBILIZED ONTO CULTURE SUBSTRATES THROUGH HEPARIN AND MATRIGEL ENHANCES DNA-SYNTHESIS IN PRIMARY RAT HEPATOCYTES
复制标题
DOI：
10.1006/excr.1994.1058
复制
发表时间：
1994-03-01
影响因子：
3.7
通讯作者：
AKAMATSU, N
中科院分区：
文献类型：
--
作者：
KATO, S;ISHII, T;AKAMATSU, N
新制备的糖胺聚糖磷脂酰乙醇胺（PE）结合物可通过疏水作用固定在固相上。原代大鼠肝细胞培养的I型胶原蛋白或层粘连蛋白基板含有肝素-PE和肝细胞生长因子（HGF）启动的DNA合成进行了测定。肝细胞对加入到培养基中的肝细胞生长因子的反应是通过活跃的DNA合成和明显的细胞扩散，而不管底物上是否存在肝素-PE。含肝素-PE的底物与HGF预孵育，但不与表皮生长因子预孵育，在培养基中不存在这些有丝分裂原的情况下显著增强DNA合成和细胞扩散。通过省略肝素-PE或用1 M NaCl洗涤HGF处理的基底来消除增强，这表明HGF通过肝素固定到基底。这些现象与层粘连蛋白基板比与I型胶原基板更明显。硫酸软骨素-PE仅部分取代肝素-PE，但Matrigel给出了与含有肝素-PE的底物类似的结果。含有肝素-PE的I型胶原和层粘连蛋白底物与各自的对照底物相比结合了更高量的HGF，而含有硫酸软骨素-PE的底物都没有。然而，HGF与底物结合的增加不如DNA合成的增加那么明显，这表明后者不仅仅是由于前者，而是由于固定化HGF和肝素的协同作用。两者合计，这些结果表明，肝细胞生长因子可以被困在细胞外基质中，可能通过硫酸乙酰肝素在体内，从而作为一个有丝分裂原的肝细胞与硫酸乙酰肝素合作。(C)1994年出版社出版。
Newly prepared phosphatidylethanolamine (PE) conjugates of glycosaminoglycans can be immobilized to solid phase through hydrophobic interaction. Primary rat hepatocytes were cultured on type I collagen or laminin substrates containing heparin-PE and assayed for DNA synthesis initiated by hepatocyte growth factor (HGF). Hepatocytes responded to HGF that had been added to culture media by active DNA synthesis and pronounced cell spreading, regardless of the presence of heparin-PE on the substrates. The preincubation with HGF, but not with epidermal growth factor, of the substrates containing heparin-PE significantly enhanced DNA synthesis and cell spreading in the absence of these mitogens in culture media. The enhancements were abolished by omitting heparin-PE or washing the HGF-treated substrates with 1 M NaCl, suggesting the immobilization of HGF to substrates through heparin. These phenomena were more evident with laminin substrates than with type I collagen substrates. Chondroitin sulfate-PE only partly substituted for heparin-PE, but Matrigel gave results similar to those of the substrates containing heparin-PE. Both type I collagen and laminin substrates containing heparin-PE bound higher amounts of HGF than the respective control substrates, whereas neither of the substrates containing chondroitin sulfate-PE did. However, the increase in HGF binding to the substrates was not as evident as the increase in DNA synthesis, suggesting that the latter is not simply due to the former but due to a concerted action of the immobilized HGF and heparin. Taken together, these results suggest that HGF can be trapped in extracellular matrix, probably through heparan sulfate in vivo, thereby acting as a mitogen for hepatocytes in cooperation with heparan sulfate. (C) 1994 Academic Press, Inc.