Fetal tissue engineering from amniotic fluid
Fetal tissue engineering from amniotic fluid
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DOI：
10.1016/s1072-7515(02)01834-3
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发表时间：
2003-04-01
影响因子：
5.2
通讯作者：
Fauza, DO
中科院分区：
文献类型：
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作者：
Kaviani, A;Guleserian, K;Fauza, DO
背景：我们最近在动物模型中表明，羊水可以成为胎儿组织工程的细胞来源。本研究旨在确定胎儿组织构建体是否也可以用人类羊水中常见的细胞进行工程改造。 研究设计：从妊娠 15 至 19 周孕妇 (n=6) 的羊水中获得的细胞在 Dulbecco's Modified Eagle's 培养基 (Sigma Chemical, St Louis, MO) 中培养，该培养基含有 20% 胎牛血清和 5 ng/mL 碱性成纤维细胞生长因子，浓度为 95% 37°C 的加湿、5% CO2 室。然后将形态不同的细胞亚群与其余细胞机械分离并选择性扩增。通过使用针对标准中间丝和表面抗原的抗体进行免疫荧光染色来确定该羊水细胞亚群的谱系。通过氧化测定测定细胞增殖率。细胞扩增后，将羊水细胞集落静态和动态接种到无纺布、1毫米厚的聚乙醇酸聚合物支架和无细胞人真皮上72小时。通过扫描电子显微镜分析所得构建体。 结果：羊水细胞平滑肌肌动蛋白、波形蛋白、细胞角蛋白 18 和成纤维细胞表面蛋白呈阳性染色，结蛋白、分化簇 31 和血管性血友病因子呈阴性染色（Dako，卡彭特里亚，加利福尼亚州）。这些发现与间充质、成纤维细胞-肌成纤维细胞谱系一致。根据增殖测定的结果，间充质羊水细胞可以在培养物中快速扩增。羊水细胞结构的扫描电子显微镜显示聚合物基质周围有致密、汇合的细胞层，细胞牢固地粘附在 PGA 和 Alloderm（Lifecell Corp，Branchburg，NJ）支架上。没有观察到细胞死亡的证据。结论：胎儿间充质细胞亚群可以从人羊水中持续分离出来，并在体外快速扩增。人间充质羊水细胞牢固地附着在聚乙醇酸聚合物和无细胞人真皮上。羊水可以成为胎儿组织工程有价值且实用的细胞来源。 (C) 2003 年，美国外科医生学会。
BACKGROUND: We have recently shown, in an animal model, that amniotic fluid can be a source of cells for fetal tissue engineering. This study was aimed at determining whether fetal tissue constructs could also be engineered from cells normally found in human amniotic fluid.STUDY DESIGN: Cells obtained from the amniotic fluid of pregnant women at 15 to 19 weeks of gestation (n=6) were cultured in Dulbecco's Modified Eagle's medium (Sigma Chemical, St Louis, MO) containing 20% fetal bovine serum and 5 ng/mL basic fibroblast growth factor in a 95% humidified, 5% CO2 chamber at 37degrees C. A subpopulation of morphologically distinct cells was then mechanically isolated-from the rest and selectively expanded. The lineage of this subpopulation of amniocytes was determined by immunofluorescent staining with antibodies against standard intermediate filaments and surface antigens. Cell proliferation rates were determined by oxidation assay. After cell expansion, colonies of amniocytes were statically and dynamically seeded onto both unwoven, 1-mm-thick polyglycolic acid polymer scaffold and acellular human dermis for 72 hours. The resulting constructs were analyzed by scanning electron microscopy.RESULTS: Amniocytes stained positively for smooth muscle actin, vimentin, cytokeratin 18, and fibroblast surface protein, and negatively for desmin, cluster of differentiation 31, and von Willebrand's factor (Dako, Carpenteria, CA). These findings are consistent with a mesenchymal, fibroblast-myofibroblast cell lineage. Mesenchymal amniocytes could be rapidly expanded in culture, based on results of the proliferation assay. Scanning electron microscopy of amniocyte constructs revealed dense, confluent layers of cells surrounding the polymer matrices and firm cell adhesion to both PGA and Alloderm (Lifecell Corp, Branchburg, NJ) scaffolds. No evidence of cell death was observed.CONCLUSIONS: Subpopulations of fetal mesenchymal cells can be consistently isolated from human amniotic fluid and rapidly expanded in vitro. Human mesenchymal amniocytes attach firmly to both polyglycolic acid polymer and acellular human dermis. The amniotic fluid can be a valuable and practical cell source for fetal tissue engineering. (C) 2003 by the American College of Surgeons.