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The effects of different concentrations of sodium selenite on the in vitro maturation of preantral follicles in serum-free and serum supplemented media.

ABSTRACT: PURPOSE: This study was to investigate the effect of sodium selenite (SS) on in vitro maturation of mouse preantral follicles. METHODS: The isolated preantral follicles were cultured in TCM 199 medium supplemented with different concentrations (0, 5, 10, 15 ng/ml) of SS and 3 mg/ml bovine serum albumin (BSA) or 5% Fetal Bovine Serum (FBS). The ovulation was induced by addition of 1.5 IU/ml human chorionic gonadotropin. The size and development of follicles and oocytes were assessed by calibrated eyepiece. RESULTS: The survival rates of follicles in FBS supplemented groups containing 5 and 10 ng/ml SS (88.23%, 90.83%) were higher than other groups (P < 0.05 and P < 0.001 respectively). The mean diameter of follicles (199.84 +/- 15.58 microm) and the percentage of MII oocyte (33.08%) were higher in FBS supplemented group containing 10 ng/ml SS (P < 0.001). CONCLUSION: The sodium selenite and FBS improve the in vitro growth and maturation of mouse preantral follicles.

SUBMITTER: Abedelahi A

PROVIDER: S-EPMC2582093 | biostudies-other | 2008 Sep-Oct

REPOSITORIES: biostudies-other

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Project description:Surgical strategies to treat articular cartilage injury such as microfracture, expose human bone marrow stem cells (hMSCs) to synovial fluid and its components. High molecular weight hyaluronan (hMwt HA) is one of the most abundant bioactive macromolecules of healthy synovial fluid (hSF) and it plays an important role in the protection of opposing articular cartilage surfaces within the synovial joint. Although hMwt HA has been extensively used to attempt the engineering of the cartilage tissue, its effect as media supplement has not been established. Indeed, current media are often simple in their composition and doesn't recapitulate the rheological and biological features of hSF. In addition, critical in vivo molecules that can potentially change the chondrogenic behavior of hBMSCs to make the in vitro results more predictive of the real in vivo outcome, are lacking. In order to be one step closer to the in vivo physiology of hSF, a new culture media supplemented with physiological level of hMwt HA was developed and the effect of the hMwt HA on the chondrogenesis of hMSCs that would be present in a traumatic defect after marrow stimulation techniques, was investigated. hBMSC-seeded fibrin-polyurethane constructs were cultured in a serum free chondropermissive control medium (HA- TGF?-). This medium was further supplemented with 10 ng/mL TGF?1 (HA- TGF?+) or 2 mg/ml hMwt HA 1.8 MDa (HA+ TGF?-) or both (HA+ TGF?+). Alternatively, 1 MDa HA was mixed with the fibrin at 0.2 mg/ml (HASc TGF?+). The effect of hMwt HA on hMSC differentiation was investigated at the gene expression level by RT-qPCR and total DNA, sulfated glycosaminoglycans and Safranin O staining were evaluated. Addition of hMwt HA to the culture media, significantly increased the synthesis of sulfated glycosaminoglycans, especially in the early days of chondrogenesis, and reduced the upregulation of the hypertrophic cartilage marker collagen X. hMwt HA added inside the fibrin gel(HASc TGF+) led to the best matrix deposition. hMwt HA can be one key medium component in a more reliable in vitro/ex vivo system to reduce in vitro artifacts, enable more accurate pre-screening of potential cartilage repair therapies and reduce the need for animal studies.

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The effects of different concentrations of sodium selenite on the in vitro maturation of preantral follicles in serum-free and serum supplemented media.

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