Lundgren E, Terracio L, Mårdh S, Borg T K
Exp Cell Res. 1985 Jun;158(2):371-81. doi: 10.1016/0014-4827(85)90462-8.
Calcium-tolerant myocytes were isolated from adult rat hearts by collagenase perfusion and plated on various substrates in serum-free medium and their adhesion to various extracellular matrix (ECM) components was determined. The myocytes attached readily to dishes coated with collagen type IV (C-IV), laminin (LN), and to fetal bovine serum (FBS) in a manner dependent on the concentration of the components. Substantially fewer myocytes adhered to dishes coated with fibronectin (FN) or to uncoated plastic dishes. Cells adhered equally well to dishes coated with C-IV, LN and FBS within 1-4 h. However, when examined after 2 weeks in culture it was found that only C-IV and LN could support survival of the attached myocytes, and when cultured on C-IV or LN the myocytes were spread and had formed a dense monolayer. The actin filaments had at this time reorganized linearly along the long axis of the cell and the myocytes contracted spontaneously. Rabbit antibodies were raised against myocyte membranes and their ability to inhibit attachment to ECM components was studied. Purified IgG inhibited attachment to C-IV, while having only a minor effect on attachment to LN. These data are compatible with the presence of a specific cell surface component(s) that interacts with ECM substrates and influences cell shape and possibly thereby influences cellular functions.
通过胶原酶灌注从成年大鼠心脏中分离出耐钙心肌细胞，并将其接种于无血清培养基中的各种底物上，测定它们对各种细胞外基质（ECM）成分的黏附情况。心肌细胞很容易黏附于包被有IV型胶原（C-IV）、层粘连蛋白（LN）的培养皿以及胎牛血清（FBS）上，其黏附方式取决于这些成分的浓度。黏附于包被有纤连蛋白（FN）的培养皿或未包被的塑料培养皿上的心肌细胞明显较少。细胞在1 - 4小时内对包被有C-IV、LN和FBS的培养皿的黏附情况相同。然而，在培养2周后检查发现，只有C-IV和LN能够支持贴壁心肌细胞的存活，并且当在C-IV或LN上培养时，心肌细胞铺展并形成致密的单层。此时，肌动蛋白丝已沿细胞长轴线性重排，心肌细胞自发收缩。制备了针对心肌细胞膜的兔抗体，并研究了其抑制与ECM成分黏附的能力。纯化的IgG抑制与C-IV的黏附，而对与LN的黏附仅有轻微影响。这些数据与存在一种特定的细胞表面成分相一致，该成分与ECM底物相互作用并影响细胞形状，进而可能影响细胞功能。
