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The experiments reported herein show the effect of porcine follicular fluid (pFF) at concentrations of 2, 4, 20, 40, 200, 400, 500, and 600 µg protein/mL on HeLa cell line viability. We found that 600, 500, and 400 µg proteins/mL of pFF from small-, medium-, and large-sized ovarian follicles showed the highest viability (141.65±26.07, 153.14±16.58, and 140.52±11.71%), which was significantly different from the control group (p<0.05), but lower than the positive control. The cell morphology of those cultured in pFF from the medium- and large-sized follicles remained normal. Cell morphology tended to be abnormal at high concentrations of pFF from small-sized ovarian follicles. Thus, we suggest the applicability of pFF from medium- and large-sized ovarian follicles at concentrations of 40, 200, 400, 500, and 600 µg protein/mL as a substitute for heat-treated fetal bovine serum (HTFBS), which is more expensive. However, it should be noted that not only did pFF extracted from small-sized ovarian follicles have no significant effect on the viability of HeLa cell lines; cell morphology tended to be abnormal at high concentrations of pFF. This study contributes to biotechnology research by discovering applications of pFF extracted from ovarian follicles of different sizes as supplements in culture media.
Adams, T., Anwar, R., Mfarej, M., Rundatz, T., Coyle, M., and Mc Laughlin, J. S. (2015). Nutritional stress of cultured Vero cells causes altered growth and morphology as seen in neoplastic transformation. American Journal of Undergraduate Research, 12(3), 63-75.
Ali, A., Coenen, K., Bousquet, D., and Sirard. M. A. (2004). Origin of bovine follicular fluid and its effect during in vitro maturation on the developmental competence of bovine oocytes. Theriogenology, 62(9), 1596-1606.
Ali, S., Ahmad, N., Akhtar, N., and Zia-Ur-Rahman, Noakes, D. E. (2008). Metabolite contents of blood serum and fluid from small and large sized follicles in dromedary camels during the peak and the low breeding seasons. Animal Reproduction Science, 108(3-4), 446-456.
Areekijseree, M., Thongpan, A., and Vejaratpimol, R. (2005). Morphological features of porcine oviductal epithelial cellsand cumulus-oocyte complex. Kasetsart Journal: Natural Science, 39(1), 136-144.
Ayoub, M. A., and Hunter, A. G. (1993). Inhibitory effect of bovine follicular fluid on in vitro maturation of bovine oocytes. Journal of Dairy Science, 76(1), 95-100.
Brunner, D., Frank, J., and Appl, H., Schöffl, H., Pfaller, G., Gstraunthaler, G., (2010). Serum-free cell culture: The serum-free media interactive online database. ALTEX, 27(1), 53-62.
Chacon, E., Acosta, D., and Lemasters, J. J. (1997). Primary cultures of cardiac myocytes as in vitro models for pharmacological and toxicological assessments. In Vitro Methods in Pharmaceutical Research, 209-223.
Ducolomb, Y., González-Márquez, H., Fierro, R., Jiménez, I., Casas, E., Flores, D., Bonilla, E., Salazar, Z., and Betancourt, M. (2013). Effect of porcine follicular fluid proteins and peptides on oocyte maturation and their subsequent effect on in vitro fertilization. Theriogenology, 79(6), 896-904.
Edwards, R. G. (1974). Follicular fluid. Journal of Reproduction and Fertility, 37(1), 189-219.
Hasasna, H. E. l., Athamneh, K., Samri, H. A. l., Karuvantevida, N., Dhaheri, Y. A. l., Hisaindee, S., Ramadan, G., Tamimi, N. A. l., AbuQamar, S., Eid, A., and Iratni, R. (2015). Rhus coriaria induces senescence and autophagic cell death in breast cancer cells through a mechanism involving p38 and ERK1/2 activation. Scientific Reports, 5, 13013.
Ito, M., Iwata, H., Kitagawa, M., Kon, Y., Kuwayama, T., and Monji, Y. (2008). Effect of follicular fluid collected from various diameter follicles on the progression of nuclear maturation and developmental competence of pig oocytes. Animal Reproduction Science, 106(3-4), 421-430.
Kimura, N., Konno, Y., Miyoshi, K., Matsumoto, H., and Sato, E. (2002). Expression of hyaluronan synthases and CD44 messenger RNAs in porcine cumulus-oocyte complexes during in vitro maturation. Biology of Reproduction, 66(3), 707-717.
Kniss, D. A., and Summerfield, T. L. (2014). Discovery of HeLa cell contamination in HES cells: Call for cell line authentication in reproductive biology research. Reproductive Sciences, 21(8), 1015-1019.
Ledwitz-Rigby, F., Rigby, B. W., Gay, V. L., Stetson, M., Young, J., and Channing, C. P. (1977). Inhibitory action of porcine follicular fluid upon granulosa cell luteinization in vitro: assay and influence of follicular maturation. Journal of Endocrinology, 74(2), 175-184.
Lowry, O., Rosebrough, N., Farr, A., and Randall, R. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193(1), 265-275.
Majumdar, S. K., Valdellon, J. A., and Brown, K. A. (2001). In vitro investigations on the toxicity and cell death induced by tamoxifen on two non-breast cancer cell types. Journal of Biomedicine and Biotechnology, 1(3), 99-107.
Mettasart, W. (2009). Cell and protein secretion from porcine oviduct and ovary in estrous cycle. (Master thesis, Silpakorn University).
Oberlender, G., Murgas, L. D. S., Zangeronimo, M. G., da Silva, A. C., de Alcantara Menezes, T., Pontelo, T. P., and Vieira, L. A. (2013). Role of insulin-like growth factor-I and follicular fluid from ovarian follicles with different diameters on porcine oocyte maturation and fertilization in vitro. Theriogenology, 80(4), 319-327.
Pongsawat, W., and Youngsabanant, M. (2019). Porcine cumulus oocyte complexes (pCOCs) as biological model for determination on in vitro cytotoxic of cadmium and copper assessment. Songklanakarin Journal of Science and Technology, 41(5),1029-1036.
Romero-Arredondo, A., and Seidel, G. E. J. (1994). Effects of bovine follicular fluid on maturation of bovine oocytes. Theriogenology, 41(2), 383-394.
Suchanek, E., Simunic, V., Juretic, D., and Grizelj, V. (1994). Follicular fluid contents of hyaluronic acid, follicle-stimulating hormone and steroids relative to the success of in vitro fertilization of human oocytes. Fertility and Sterility, 62(2), 347-352.
Youngsabanant-Areekijseree, M., Tungkasen, H., Srinark, C., and Chuen-Im, T. (2019). Determination of porcine oocyte and follicular fluid proteins from small, medium, and large follicles for cell biotechnology research. Songklanakarin Journal Science Technology, 41(1), 192-198.
Youngsabanant, M., Gumlungpat, N., and Panyarachun, B., (2019). In vitro characterization and viability of Vero cell lines supplemented with porcine follicular fluid proteins study. Science, Engineering and Health Studies, 13(3), 143-152.