| Abstrakt: |
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. [ABSTRACT FROM AUTHOR] |
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