Bone marrow mesenchymal stem cell transplantation improves ovarian function and structure in rats with chemotherapy-induced ovarian damage
Introduction
Chemotherapy has been used widely in the treatment of various malignancies and autoimmune diseases, and recent advances in chemotherapy have resulted in significant long-term survival improvements for these patients [1]. However, patients undergoing chemotherapy have to face some severe adverse effects and, in female patients, chemotherapy-induced irreversible damage to the ovarian tissues remains of great concern. Hot flashes, osteoporosis, sexual dysfunction and infertility are all the consequences of premature ovarian failure (POF) induced by chemotherapy [2., 3., 4.].
Chemotherapy-induced ovarian damage is directly correlated with the dose, kind of therapeutic agents used, duration of therapy and age of the patient undergoing treatment [5]. Among the commonly used chemotherapeutic medications, alkylating agents are shown to be the most gonadotoxic. Recent studies both in vitro and in vivo have proved that these agents can induce abnormal apoptosis of oocytes and granulosa cells (GC) and consequently loss of follicles [6., 7., 8.].
Mesenchymal stem cells (MSC) residing within the bone marrow (BM) microenvironment are pluripotent adult stem cells, whose multipotency, easy isolation and culture as well as high ex vivo expansive potential make them attractive candidates for stem cell therapies [9]. Successful attempts of BM-derived MSC transplantation for repairing spinal, cardiac and cutaneous injuries, for instance, have been reported [10, 11]. MSC can differentiate into osteoblasts, chondrocytes, adipocytes, cardiocytes, neural cells and hematopoietic-supporting stroma, and may therefore partly replace the impaired cells [12]. The mechanisms of repair effects also involves their supplying cytokines for angiogenesis, anti-apoptosis and mitogenesis, such as vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), insulin-like growth factor-1 (IGF-1) and basic fibroblast growth factor (b-FGF) [13]. However, currently little is known about the therapeutic potential of MSC for chemotherapy-induced ovarian damage. We conducted this study to investigate the following topics: whether (1) MSC inhibit chemotherapy-induced apoptosis of ovarian GC in vitro; (2) MSC ameliorate the structural and functional damage of ovaries as a result of chemotherapy in a rat model; and (3) the therapeutic effects of MSC are mediated by secreted anti-apoptotic cytokines including VEGF, IGF-1 and HGF.
Section snippets
Experimental animals
All animal procedures were conducted in accordance with the institutional guidelines of the Southern Medical University (Guangzhou, P. R. China) for the care and use of laboratory animals. Adult female Wistar rats, weighing 180–210 g, were fed rat chow and water ad libitum under controlled temperature (30 ±; 2°C) and light (14 h light, 10 h dark) conditions. Vaginal smears were obtained daily. Only those showing at least two consecutive normal 4-day vaginal estrus cycles were included in the
Characterization and labeling of cultured MSC
Cells of the third passage of MSC in culture were adherent and morphologically resembled fibroblasts (Figure 1A). Most of the cultured MSC expressed CD29 and CD44 but were negative for markers of the hematopoietic lineage, including CD34 and CD45, consistent with previously published data on MSC surface markers [16] (Figure 1C).
A MOI of 100 appeared to be optimal for Ad-GFP transfection of MSC, resulting in a transfection efficiency of 95%. The transfected MSC showed bright green fluorescence
Discussion
MSC have attracted interest for their possible use for both cell and gene therapies because of their capacity for self-renewal and multipotentiality for differentiation [17]. These cells can be induced, either in vitro or in vivo, to differentiate terminally into osteoblasts, chondrocytes, adipocytes, tenocytes, cardiomyoctyes, neural cells and hematopoietic-supporting stroma. Transplantation of MSC directly into adult rat brain and heart reduces functional deficits resulting from stroke [18]
Acknowledgements
This work was supported by a grant from the Science and Technology Project of Guangdong Province (2006B35901003).
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