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Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture

机译:使用可生物吸收的聚(L-乳酸)/聚(羟基丁酸酯-共羟基戊酸酯)的混合物作为Vero细胞培养的表面

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摘要

Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37°C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.
机译:Vero细胞是一种从非洲绿猴(Cercopithecus aethiops)肾脏建立的细胞系,于37°C的F-10 Ham培养基中添加10%胎牛血清,并在聚L-乳酸膜上培养( PLLA),聚(羟基丁酸酯-共羟基戊酸酯)(PHBV)及其混合物的比例不同(100 / 0、60 / 40、50 / 50、40 / 60和0/100)。本研究通过扫描电子显微镜评估了培养24小时后在不同聚合物基质上生长的细胞的形态。接种2小时后还分析细胞粘附。为了进行细胞生长评估,将细胞保持培养48、120、240和360小时。为了进行细胞化学研究,将细胞培养120或240小时,固定,处理以进行组织学分析,并用pH 4.0的甲苯胺蓝和pH 2.5的二甲苯啶丽春红染色。我们的结果表明,使用60/40和50/50的混合物时,细胞粘附性更好,尽管在所有测试的混合物中细胞都能够生长和增殖。使用PLLA / PHBV(50/50)时,在多孔和光滑区域观察到略微扁平的细胞。 PLLA / PHBV(40/60)共混物在光滑区域呈现出扁平化的细胞。没有毛孔的PLLA / PHBV(0/100)也支持由细丝相互连接的散布细胞。组织学切片显示,细胞在不同基质上以汇合的单层生长。细胞化学分析显示嗜碱性细胞,表明大量的RNA和蛋白质。因此,我们检测到由于混合比例的改变而引起的细胞形态变化。这表明当在各种PLLA / PHBV混合表面上时,细胞会改变其分化模式。

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