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首页> 外文期刊>Reproduction in Domestic Animals >Effects of Prolonged in vitro Culture and Cryopreservation on Viability, DNA Fragmentation, Chromosome Stability and Ultrastructure of Bovine Cells from Amniotic Fluid and Umbilical Cord
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Effects of Prolonged in vitro Culture and Cryopreservation on Viability, DNA Fragmentation, Chromosome Stability and Ultrastructure of Bovine Cells from Amniotic Fluid and Umbilical Cord

机译：长时间体外培养和冷冻保存对羊水和脐带牛细胞活力，DNA片段化，染色体稳定性和超微结构的影响

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Contents The objective of this work was to study cellular types that did not participated in the gastrulation process, amniotic fluid cells (AFCs) and umbilical cord cells (UCCs), in conditions of long-term culture and cryopreserved with different solutions. The AFCs and UCCs were used in a comparative study with ear fibroblast cells (EFCs) that were cultured in vitro until 20 cellular passages and cryopreserved in 10% dimethylsulphoxide (DMSO), 5% dimethyl formamide (DMF) and 7% glycerol (Gly) solutions. The cellular viability, ultrastructure, DNA fragmentation and chromosome stability were evaluated to determine the cellular type most resistant. In all cell types, it was possible to evaluate the AFCs until 15 passages and UCCs until 20 passages with different periods of cellular growth to reach the confluence phase. Solutions containing 10% DMSO ensured viability of 90.33 +/- 5.58%, 90.56 +/- 4.40% and 81.90 +/- 3.31%, respectively for EFCs, AFCs and UCCs, being significantly more efficient and with less variation than other cryoprotectant solutions. The AFCs were more sensitive to cryopreservation and presented low viability rate at the passage 20 (17.2 +/- 8.87%). There was no change in karyotype and nuclear fragmentation was low in all cellular passages studied. With the scanning electron analysis was possible the characterization of AFCs and UCCs in suspension. The three cellular types of cells presented different shapes and characteristics on the surface. The results demonstrate that bovine AFCs and UCCs can be isolated, cultured in vitro and cryopreserved in 10% DMSO, not causing damage to DNA and chromosomes. The UCCs were more resistant than AFCs in all aspects
机译：内容这项工作的目的是研究在长期培养条件下并用不同溶液冷冻保存的未参与胃泌尿过程的细胞类型，羊水细胞（AFC）和脐带细胞（UCC）。将AFC和UCC与耳成纤维细胞（EFC）进行对比研究，将其体外培养直至20次细胞传代，并冷冻保存在10％的二甲基亚砜（DMSO），5％的二甲基甲酰胺（DMF）和7％的甘油（Gly）中解决方案。评价细胞的生存力，超微结构，DNA片段化和染色体稳定性，以确定最有抗性的细胞类型。在所有细胞类型中，有可能评估AFC直到15代，而UCC直到20代，具有不同的细胞生长周期才能达到融合阶段。含有10％DMSO的溶液分别确保EFC，AFC和UCC的生存力分别为90.33 +/- 5.58％，90.56 +/- 4.40％和81.90 +/- 3.31％，与其他防冻剂溶液相比，其效率显着提高且变化较小。 AFC对冷冻保存更敏感，在20代时存活率较低（17.2 +/- 8.87％）。在所有研究的细胞传代中，核型无变化，核碎裂率低。通过扫描电子分析，可以对悬浮液中的AFC和UCC进行表征。三种细胞类型的细胞在表面上呈现出不同的形状和特征。结果表明，牛AFC和UCC可以分离，体外培养，并在10％DMSO中冷冻保存，不会对DNA和染色体造成破坏。在所有方面，UCC比AFC更耐

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