Abstract

Objective. Amniotic fluid contains a broad range of cells called amniocytes. These cells exhibit various morphologies, activities and features, depending on gestational age and fetal development. Here we present comprehensive data on growth properties of different subpopulations from single clone to senescence under different culture media. Materials and methods. Amniotic fluid samples were obtained from high risk pregnant women, who were candidates for amniocentesis. We investigated samples from 68 cases after completion of their diagnosis procedure. Accordingly, amniotic fluid cells were cultured until cells became senescent and culture growth ceased. Cells were cultured by three protocols: (1) AmnioMAX-II supplemented with 20 mM HEPES and 1% PenStrep; (2) DMEM supplemented with 4 mM L-glutamine, 10 mM HEPES, 15% FBS and 1% PenStrep; (3) a modified medium composed of 2 : 1 v/v DMEM : AmnioMAX-II which were supplemented as indicated in the second and first protocols, respectively. Culture characteristics and growth kinetics of different cell populations were further investigated. Results. All amniocytes had the ability to form colonies due to their fetal origin. Here, we show that life span of various cell types in amniotic fluid differs significantly: F-type cells have longer life span (on average 15 passages) compared to AF-type cells (6.6 on average) and E-type cells (4.6 on average). Student’s t-test analysis revealed a non-significant difference between the life span of amniocytes in AmnioMAX-II and DMEM-AmnioMAX-II (2 : 1 v/v), nearly 7 passages on average, which was significantly higher than their life span in DMEM (4 passages on average). Conclusion. Our study showed that all three cell types are clonogenic with distinguishable characteristics. Considering the effects of medium on growth properties of these subpopulations, we have found that morphological features and growth characteristics of amniocytes subjected to in vitro expansion can be modulated by the medium formulation used for cultivation.

中文翻译：

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人类羊细胞：从单个克隆到衰老的亚群的形态，频率和生长特性的综合研究

摘要

目的。羊水含有多种称为羊水的细胞。这些细胞表现出各种形态，活动和特征，具体取决于胎龄和胎儿发育。在这里，我们介绍了在不同培养基下从单个克隆到衰老的不同亚群的生长特性的综合数据。材料和方法。羊水样本是从高危孕妇中抽取的，她们是羊膜穿刺术的候选人。在完成诊断程序后，我们调查了68例患者的样本。因此，培养羊水细胞直到细胞衰老并且培养物生长停止。通过三种方案培养细胞：（1）补充了20 mM HEPES和1％PenStrep的AmnioMAX-II；（2）补充4 mM L-谷氨酰胺，10 mM HEPES，15％FBS和1％PenStrep的DMEM；（3）由2∶1 v / v DMEM：AmnioMAX-II组成的改良培养基，分别按照第二和第一方案中的指示进行补充。进一步研究了不同细胞群体的培养特性和生长动力学。结果。由于它们的胎儿来源，所有羊膜细胞都具有形成菌落的能力。在这里，我们发现羊水中各种细胞类型的寿命存在显着差异：与AF型细胞（平均6.6）和E型细胞（4.6）相比，F型细胞的寿命更长（平均15次传代）。平均）。学生的t检验分析显示，AmnioMAX-II和DMEM-AmnioMAX-II中羊水细胞的寿命之间没有显着差异（2：1 v / v），平均传代近7次，显着高于它们的寿命在DMEM中（平均4遍）。结论。我们的研究表明，所有三种细胞类型都是具有明显特征的克隆性。考虑到培养基对这些亚群的生长特性的影响，我们发现，体外培养的羊膜细胞的形态特征和生长特性可以通过用于培养的培养基配方来调节。