Endothelial cells modulate the proliferation of mural cell precursors via platelet-derived growth factor-BB and heterotypic cell contact
Endothelial cells modulate the proliferation of mural cell precursors via platelet-derived growth factor-BB and heterotypic cell contact
复制标题
DOI：
10.1161/01.res.84.3.298
复制
发表时间：
1999-02-19
影响因子：
20.1
通讯作者：
D'Amore, PA
中科院分区：
文献类型：
--
作者：
Hirschi, KK;Rohovsky, SA;D'Amore, PA
胚胎学数据表明，内皮细胞（ECs）直接招募和壁细胞前体的分化。我们已经开发了体外共培养系统来模拟这些事件中的一些，并已表明，内皮细胞直接迁移的未分化的间充质细胞（10 T1/2细胞），并诱导其向平滑肌细胞/周细胞谱系分化。本研究采用琼脂糖凝胶电泳法观察内皮细胞与10 T1/2细胞在无接触条件下的共培养情况。10 T1/2细胞对内皮细胞的反应中溴脱氧尿苷标记增加2倍，这被血小板衍生生长因子（PDGF）-B的中和抗血清完全抑制。抗PDGF-A、碱性成纤维细胞生长因子或转化生长因子（TGF）-β的抗血清对内皮细胞刺激的10 T1/2细胞增殖没有影响。EC增殖不受10 T1/2细胞在无接触的情况下共培养的影响。然后将细胞共培养，以便允许接触。双标记和荧光激活细胞分选仪分析显示，EC和10 T1/2细胞的生长抑制分别为43%和47%。接触EC-10 T1/2细胞共培养物的条件培养基分别抑制两种细胞类型的生长61%和48%。尽管我们先前已经显示了TGF-β在共培养诱导的壁细胞分化中的作用，但是由接触共培养物或条件培养基引起的生长抑制并不被针对TGF-β的中和抗血清的存在所抑制。直接共培养中10 T1/2细胞增殖的降低不能归因于EC中PDGF-B或10 T1/2细胞中PDGF受体β的下调。2个细胞。我们的数据表明，在EC招聘间充质细胞和异型细胞-细胞接触和可溶性因子在血管组装过程中的生长控制中发挥作用的增殖发生调制。
Embryological data suggest that endothelial cells (ECs) direct the recruitment and differentiation of mural cell precursors. We have developed in vitro coculture systems to model some of these events and have shown that ECs direct the migration of undifferentiated mesenchymal cells (10T1/2 cells) and induce their differentiation toward a smooth muscle cell/pericyte lineage. The present study was undertaken to investigate cell proliferation in these cocultures, ECs and 10T1/2 cells were cocultured in an underagarose assay in the absence of contact. There was a 2-fold increase in bromodeoxyuridine labeling of 10T1/2 cells in response to ECs, which was completely inhibited by the inclusion of neutralizing antiserum against platelet-derived growth factor (PDGF)-B, Antisera against PDGF-A, basic fibroblast growth factor, or transforming growth factor (TGF)-beta had no effect on EC-stimulated 10T1/2 cell proliferation. EC proliferation was not influenced by coculture with 10T1/2 cells in the absence of contact. The cells were then cocultured so that contact was permitted. Double labeling and fluorescence-activated cell sorter analysis revealed that ECs and 10T1/2 cells were growth-inhibited by 43% and 47%, respectively. Conditioned media from contacting EC-10T1/2 cell cocultures inhibited the growth of both cell types by 61% and 48%, respectively. Although we have previously shown a role for TGF-beta in coculture-induced mural cell differentiation, growth inhibition resulting from contacting cocultures or conditioned media was not suppressed by the presence of neutralizing antiserum against TGF-beta, Furthermore, the decreased proliferation of 10T1/2 cells in the direct cocultures could not be attributed to downregulation of the PDGF-B in ECs or the PDGF receptor-beta in the 10T1/2 cells. Our data suggest that modulation of proliferation occurs during EC recruitment of mesenchymal cells and that heterotypic cell-cell contact and soluble factors play a role in growth control during vessel assembly.