Growth factors and their receptors derived from human amniotic cells in vitro
Growth factors and their receptors derived from human amniotic cells in vitro
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DOI：
10.5603/fhc.2014.0019
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发表时间：
2014-01-01
影响因子：
1.5
通讯作者：
Kawiak, Jerzy
中科院分区：
文献类型：
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作者：
Grzywocz, Zofia;Pius-Sadowska, Ewa;Kawiak, Jerzy
体外研究表明，p53产生的生长因子参与血管生成、上皮再生和免疫调节。本研究的目的是研究人羊膜组织和羊膜细胞产生的生长因子及其受体。分离人羊膜（hAM），并解剖羊膜环用于体外分析。通过使用不同的方法消化一些羊膜片段以获得两种细胞级分，分析其间充质和上皮细胞标记物。羊膜环和人羊膜细胞组分在无蛋白质培养基中培养。用人生长因子抗体阵列分析分泌到培养基中的蛋白质。将条件培养基加入人脐静脉上皮细胞（HUVEC）中，以测试迁移（划痕试验）和增殖（Ki 67表达）的刺激。部分1细胞同时表达细胞角蛋白和间充质细胞标志物，表明其由人羊膜上皮细胞（hAEC）和间充质基质细胞（hAMSC）的混合物组成。部分2细胞主要表达细胞角蛋白，因此被设计为hAEC。培养细胞的蛋白质分泌随时间增加。hAM培养物分泌EGF-R、IGF和IGFBP-2、-3和-6;细胞级分1分泌NT-4，而细胞级分2分泌G-CSF、M-CSF和PDGF。人脐静脉内皮细胞条件培养液刺激HUVECs迁移。我们首次发现人羊膜细胞和羊膜细胞分泌IGFBP-6、MCSF-R、PDGF-AB、FGF-6、IGFBP-4、NT-4和VEGF-R3。我们发现，细胞组分1，细胞组分2，和整个羊膜分泌不同的蛋白质，可能是由于不同比例的羊膜衍生细胞和不同的细胞-细胞相互作用。hAM细胞因子在体外保持功能并诱导HUVECs迁移增强。在羊膜或羊膜细胞培养基中发现的生长因子和受体可能用于再生医学。
In vitro studies have shown that amnion-produced growth factors participated in angiogenesis, re-epithelialization, and immunomodulation. The aim of our study was to investigate the growth factors and receptors produced by human amnion tissue and amniotic cells. Human amnions (hAM) were isolated, and amnion circles were dissected for in vitro analysis. Some amnion fragments were digested by the use of different methods to obtain two cell fractions, which were analysed for mesenchymal and epithelial cell markers. Amniotic circles and human amniotic cell fractions were cultured in a protein-free medium. Proteins secreted into the culture medium were analysed with a human growth factor antibody array. Conditioned culture media were added to human umbilical vein epithelial cells (HUVECs) to test for stimulation of migration (scratch test) and proliferation (Ki67 expression). Fraction 1 cells expressed both cytokeratin and mesenchymal cell markers which indicated that it was composed of a mixture of human amnion epithelial cells (hAECs) and mesenchymal stromal cells (hAMSCs). Fraction 2 cells mainly expressed cytokeratin and, therefore, were designed as hAECs. Secretion of proteins by the cultured cells increased with time. The hAM cultures secreted EGF-R, IGF, and IGFBP-2, -3 and -6; Cell Fraction 1 secreted NT-4, whereas Cell Fraction 2 secreted G-CSF, M-CSF, and PDGF. Conditioned media of hAM cultures stimulated HUVECs migration. We have showed for the first time that human amnions and amniotic cells secreted IGFBP-6, MCSF-R, PDGF-AB, FGF-6, IGFBP-4, NT-4, and VEGF-R3. We found that Cell Fraction 1, Cell Fraction 2, and the whole amnion secreted different proteins, possibly due to different proportions of amnion-derived cells and different cell-cell interactions. The hAM cell factors remained functional in vitro and induced intensified migration of HUVECs. The growth factors and receptors found in amnion or amniotic cell media might be used for regenerative medicine.