Induction of neovascularization in vivo and endothelial proliferation in vitro by tumor-associated macrophages.
Induction of neovascularization in vivo and endothelial proliferation in vitro by tumor-associated macrophages.
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发表时间：
1984-12
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通讯作者：
P. Polverini;S. Leibovich
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作者：
P. Polverini;S. Leibovich
通过检查肿瘤相关巨噬细胞（TAM）及其条件培养基诱导同系大鼠角膜新血管形成和培养物中牛主动脉内皮细胞增殖的能力，研究了巨噬细胞在肿瘤新血管形成中的作用。通过酶解法从 F344 雄性大鼠中繁殖的 3-甲基胆蒽诱导的纤维肉瘤中分离 TAM，并通过连续 Percoll 密度梯度离心纯化，然后粘附到纤连蛋白包被的真皮胶原凝胶上。比较了 (a) TAM 及其 72 小时条件培养基、(b) 全肿瘤细胞悬浮液 (WTCS)、(c) 去除 TAM 的肿瘤细胞悬浮液 (TCS) 和 (d) 用 TAM 重建的去除巨噬细胞的肿瘤细胞悬浮液 (TCS + TAM) 的血管生成潜力。细胞直接注射；将条件培养基浓缩 10 倍，掺入缓释 Hydron 颗粒中，并植入角膜内。在 TAM 条件培养基培养物中测定 TAM 对牛主动脉内皮细胞生长的刺激作用，并与来自蛋白胨引发的大鼠腹膜渗出巨噬细胞的条件培养基诱导的反应进行比较。 TAM 及其条件培养基分别在 40 个角膜中的 38 个（95%）和 17 个角膜中的 15 个（88%）中诱导新生血管形成。最大血管向内生长发生在植入第五天。 WTCS（26 个角膜中的 24 个，92%）和 TCS（24 个角膜中的 17 个，71%）诱导的新生血管反应分别发生在第 7 天和第 10 天。 TCS + TAM 诱导的新生血管反应与 WTCS 引起的新生血管反应相当（20 个角膜中的 19 个，95%）。在牛主动脉内皮细胞培养物中添加 TAM 条件培养基可在 10 天内刺激细胞数量增加 10 倍。这种生长刺激作用相当于或大于来自大鼠腹膜巨噬细胞的条件培养基诱导的反应。我们的结果表明，TAM 是新血管形成和内皮细胞增殖的有效刺激剂，并且肿瘤细胞悬浮液中巨噬细胞的消耗显着降低了其血管生成潜力。这表明该肿瘤的新血管形成部分是由巨噬细胞介导的。
The role of macrophages in neovascularization of tumors was investigated by examining the ability of tumor-associated macrophages (TAM) and their conditioned culture media to induce neovascularization in the cornea of syngeneic rats and proliferation of bovine aortic endothelial cells in culture. TAM were isolated from a 3-methycholanthrene-induced fibrosarcoma propagated in F344 male rats by enzymatic dissociation and were purified by centrifugation through continuous Percoll density gradients, followed by adherence to fibronectin-coated dermal collagen gels. The angiogenic potential of (a) TAM and their 72-hour conditioned culture media, (b) whole tumor cell suspensions (WTCS), (c) tumor cell suspensions depleted of TAM (TCS), and (d) macrophage-depleted tumor cell suspensions reconstituted with TAM (TCS + TAM) were compared. Cells were injected directly; conditioned media were concentrated 10-fold, incorporated into slow-release Hydron pellets, and implanted intracorneally. Stimulation of bovine aortic endothelial cell growth by TAM was assayed in culture with TAM-conditioned media and compared with responses induced by conditioned media from peptone-elicited rat peritoneal exudate macrophages. TAM and their conditioned media induced neovascularization in 38 of 40 corneas (95%) and 15 of 17 corneas (88%), respectively. Maximal vessel ingrowth occurred by the 5th day of implantation. Neovascular responses induced by WTCS (24 of 26 corneas, 92%) and TCS (17 of 24 corneas, 71%) occurred on the 7th and 10th day, respectively. TCS + TAM induced neovascular responses comparable to those elicited by WTCS (19 of 20 corneas, 95%). Addition of TAM-conditioned media to bovine aortic endothelial cell cultures stimulated a 10-fold increase in cell number within 10 days. This growth stimulatory effect was comparable to or greater than responses induced by conditioned media from rat peritoneal macrophages. Our results demonstrate that TAM are potent stimulators of neovascularization and endothelial cell proliferation and that depletion of macrophages from tumor cell suspensions significantly decreased their angiogenic potential. This suggests that neovascularization of this tumor is mediated in part by macrophages.