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长期储存后自体血清滴眼液中生长因子的稳定性

Stability of Growth Factors in Autologous Serum Eyedrops After Long-Term Storage.

作者信息

López-García José Santiago, García-Lozano Isabel, Rivas Luis, Ramírez Nuria, Méndez Maria T, Raposo Rafaela

机构信息

a Ophthalmology Service, Hospital Cruz Roja , Madrid , Spain .

b Dry Eye Unit, Research Laboratory , Ophthalmology Service, Hospital Ramón y Cajal , Madrid , Spain .

出版信息

Curr Eye Res. 2016;41(3):292-8. doi: 10.3109/02713683.2015.1016180. Epub 2015 Mar 24.

Abstract

PURPOSE

The purpose of this study is to assess the stability of the growth factors (GF) in autologous serum eyedrops under different storage conditions.

METHODS

The concentration of epidermal growth factor (EGF), transforming growth factor-β (TGF-β1), platelet-derived growth factor AB (PDGF-AB), and albumin was measured in fresh and defrosted samples of autologous serum under different storage conditions. The fresh and defrosted samples were cooled at 4 °C, and they were studied immediately after preparation, or after defrosting, and after 1, 2, 3, and 4 weeks. The concentration of GF was also assessed after 1, 3, 6, and 9 months at -20 °C. We also investigated how the different storage conditions influence the biological effects of autologous serum on conjunctival and corneal cell cultures.

RESULTS

The concentration of EGF, TGF-β1, PDGF-AB, and albumin remained stable over the 4 weeks at 4 °C, both in fresh and in defrosted samples. Likewise, no statistically significant differences were found between the GF concentration in fresh samples and after 1, 3, 6, and 9 months of freezing at -20 °C. Moreover, no differences were found on the cell proliferation and differentiation between cultured cells with fresh or defrosted samples after 4 weeks at 4 °C or after 1, 3, 6, or 9 months at -20 °C.

CONCLUSIONS

Long-term storage of autologous serum eyedrops at -20 °C does not affect the concentration of GF, simplifies clinical logistics, and reduces the frequency of blood extractions from the patients.

摘要

目的

本研究旨在评估自体血清滴眼液中生长因子(GF)在不同储存条件下的稳定性。

方法

在不同储存条件下,对新鲜和解冻后的自体血清样本中表皮生长因子(EGF)、转化生长因子-β(TGF-β1)、血小板衍生生长因子AB(PDGF-AB)和白蛋白的浓度进行测量。新鲜和解冻后的样本在4℃下冷藏,在制备后、解冻后、1、2、3和4周后进行研究。在-20℃下储存1、3、6和9个月后也评估了GF的浓度。我们还研究了不同储存条件如何影响自体血清对结膜和角膜细胞培养的生物学效应。

结果

在4℃下,新鲜样本和解冻样本中的EGF、TGF-β1、PDGF-AB和白蛋白浓度在4周内均保持稳定。同样,新鲜样本与在-20℃下冷冻1、3、6和9个月后的GF浓度之间未发现统计学上的显著差异。此外,在4℃下保存4周或在-20℃下保存1、3、6或9个月后,用新鲜或解冻样本培养的细胞在细胞增殖和分化方面没有差异。

结论

自体血清滴眼液在-20℃下长期储存不会影响GF的浓度,简化了临床后勤工作,并减少了从患者身上采血的频率。

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