First step in developing a 3D biodegradable fibrin scaffold for an artificial ovary

Journal of Ovarian Research

Christiani A Amorim⁴  Jacques Donnez¹  Sarah R Scalercio²  Julie Vanacker⁴  Marie-Madeleine Dolmans³  Valérie Luyckx⁴ 

[1] SRI (Society for Research into Infertility), Brussels, Belgium;National Primate Centre, Secretary of Health Policy, Ministry of Health, Belém, Brazil;Gynecology Department, Cliniques Universitaires Saint-Luc, Brussels, Belgium;Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium

关键词: Scanning electron microscopy;    In vitro culture;    Ovarian stromal cells;    Artificial ovary;    Tissue engineering;    Fibrin;   

Others  :  804927
DOI  :  10.1186/1757-2215-6-83

 received in 2013-09-10, accepted in 2013-11-18,  发布年份 2013

【 摘 要 】

Background

Although transplantation of cryopreserved ovarian tissue is a promising approach to restore fertility in cancer patients, it is not advisable for women at risk of ovarian involvement due to the threat of reintroducing malignant cells. The aim of this study was therefore to find an alternative for these patients by development of an artificial ovary.

Methods

For construction of the artificial ovary matrix, we used a central composite design to investigate nine combinations of fibrinogen (mg/ml) and thrombin (IU/mL) (F/T): F1/T4, F12.5/T1, F12.5/T20, F25/T0.1, F25/T4, F25/T500, F50/T1, F50/T20 and F100/T4. From the first qualitative analyses (handling and matrix size), five combinations (F12.5/T1, F25/T4, F50/T20, F50/T1 and F100/T4) yielded positive results. They were further evaluated in order to assess fibrin matrix degradation and homogeneous cell encapsulation (density), survival and proliferation (Ki67), and atresia (TUNEL) before and after 7 days of in vitro culture. To determine the best compromise between maximizing the dynamic density (Y1) and minimizing the apoptosis rate (Y2), we used the desirability function approach.

Results

Two combinations (F12.5/T1 and F25/T4) showed greater distribution of cells before in vitro culture, reproducible degradation of the fibrin network and adequate support for isolated human ovarian stromal cells, with a high proportion of Ki67-positive cells. SEM analysis revealed a network of fibers with regular pores and healthy stromal cells after in vitro culture with both F/T combinations.

Conclusion

This study reports two optimal F/T combinations that allow survival and proliferation of isolated human ovarian cells. Further studies are required to determine if such a scaffold will also be a suitable environment for isolated ovarian follicles.

【 授权许可】

   
2013 Luyckx et al.; licensee BioMed Central Ltd.

【 图 表 】

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