The purpose of this study was to investigate the effect of estrus synchronization to altrenogest regumate (progesterone), PMSG/hCG, and artificial insemination (AI) on galectin-3, FGF-9 gene and protein expression. The morpho-metrical parameters of the endometrium and the number of corpora lutea (CL) were recorded. RNA was isolated from endometrial, oviduct and ovary tissues of non-synchronized (Control; n = 7) and AI synchronized (regumate, PMSG/hCG; n = 7) sows. The total number of CL was higher (P<0.05) in pigs treated with regumate/PMSG/hCG. The content of gelactin-3 and FGF-9 mRNA in pre-embryonic development stages increased on particular days, in control and studied in regumate/PMSG/hCG administered pigs. Gelactin-3 and FGF-9 were affected by regumate/PMSG/hCG treatment in the both pre-embryonic development stages (P<0.001, P<0.05) and encdometrial tissue (P<0.001, P<0.01). The regumate/PMSG/hCG treatment resulted in elevated expression of gelactin-3 (P<0.001) and FGF-9 (P<0.005) in oviduct and ovary tissues in comparison to control sows. Moreover, oviduct amount of gelectin-3 mRNA was higher in regumate/PMSG/hCG sows in comparison to the control group (P<0.05), whereas, expression characteristics of gelactin-3 and FGF-9 were investigated by hematoxylin and eosin stained and immunohistochemical staining. The results showed that galectin-3 and FGF-9 were significantly shown in the endometrium, oviduct and ovary tissues of the regumate/PMSG/ hCG. Presented data show that exogenous hormones administration can affect gene and protein expression in the sow reproductive tract.

The development of humanized culture system of human embryonic stem cells (hESCs) hold promise for therapeutic applications. However, conventional culture system contain animal-derived components such as fetal bovine serum and mouse embryonic fibroblasts that bear a risk of transmitting non-human pathogens and incorporation of non-human immunogenic molecules to hESCs. In this study, we developed an efficient xeno-free hESCs culture system using humanized materials, the CELLstartTM, human foreskin feeder and xeno-free medium containing knockOutTM SR XenoFree (XF-medium) without animal-derived material. The hESCs were gradually adapted to the XF-medium; 25:75, 50:50, 75:25 and 100:0. Two karyotypically normal hESC lines, SNUhES4 and H1, were used for the experiments of xeno-free culture condition. The attachment rates at xeno-free culture system were 52.6±12.4%, 67.0±16.6%, 59.0±13.9%, 28.3±2.9% in SNUhES4, 79.3±5.4%, 53.8±20.9%, 69.4 ±6.4%, 59.8±12.6% in H1 and the spontaneous differentiation rates were 42.2±12.7%, 31.4±2.9%, 40.8±14.5%, 55.2±35.5% in SNUhES4, 35.6±8.5%, 36.4±13.5%, 48.4±7.8%, 80.1±6.0% in H1 in the first four passage. Although the attachment rates were low and the spontaneous differentiation rates were high compared to that of conventional system in the early passages using this humanized culture condition, hESCs in this culture condition were found to maintain hESC characterizations; morphology, expression of cell surface markers and stable karyotype. Our results indicate that simplified compositions of humanized culture system can be applicable to the further optimization for a xeno-free culture of hESCs without the loss of pluripotency and contamination from xenogenic sources.