Factors Affecting Primary Cultures of Haliotis discus hannai Ovary-dissociated Cells and General Culture Aspects
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We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ≥10 weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture—18.4 and 19.5 days for first and second subcultures, respectively—and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.
Abstract
We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ≥10 weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture—18.4 and 19.5 days for first and second subcultures, respectively—and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.
참고문헌 (38)
- Andrae J, "Role of platelet-derived growth factors in physiology and medicine", Genes Dev, 22, pp.1276~1312 (2008)
- Auzoux-Bordenave S, "CGRP regulates the activity of mantle cells and hemocytes in abalone primary cell cultures(Haliotis tuberculata)", J Shellfish Res, 26, pp.887~894 (2007)
- Awaji M, "Gonad formation, sex differentiation and gonad maturation processes in artificially produced juveniles of the abalone, Haliotis discus hannai", Aquaculture, 239, pp.397~411 (2004)
- Chen SN, "Establishment of cell lines derived from oyster, Crassostrea gigas Thunberg and hard clam, Meretrix lusoria Röding", Meth Cell Sci, 21, pp.183~192 (1999)
- Choi SS, "Beneficial effect of silkworm hemolymph on a CHO cell system : Inhibition of apoptosis and increase of EPO production", Biotechnol Bioeng, 91, pp.793~800 (2005)
- d’Agostino RB, "Tutorial in biostatistics : propensity score methods for bias reduction in the comparison of a treatment to a nonrandomized control group", Stat Med, 17, pp.2265~2281 (1998)
- Domart-Coulon I, "Identification of media supplements that improve the viability of primary cell cultures of Crassostrea gigas oysters", Cytotechnology, 16, pp.109~120 (1994)
- Farcy E, "Identification of cDNAs encoding HSP70 and HSP90 in the abalone Haliotis tuberculata : Transcriptional induction in response to thermal stress in hemocyte primary culture", Comp Biochem Physiol B Biochem Mol Biol, 146, pp.540~550 (2007)
- Freshney RI, "Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications" (2010), Wiley-Blackwell
- Gaume B, "In vitro effects of triclosan and methyl-triclosan on the marine gastropod Haliotis tuberculate", Comp Biochem Physiol C Toxicol Pharmacol, 156, pp.87~94 (2012)
- Ghirardi M, "Aplysia hemolymph promotes neurite outgrowth and synaptogenesis of identified Helix neurons in cell culture", Invert Neurosci, 2, pp.41~49 (1996)
- Guan K, "Pluripotency of spermatogonial stem cells from adult mouse testis", Nature, 440, pp.1199~1203 (2006)
- Hyland C, "Regeneration of Aplysia bag cell neurons is synergistically enhanced by substrate-bound hemolymph proteins and laminin", Sci Rep, 4, pp.4617~ (2014)
- Khurad AM, "A new continuous cell line from larval ovaries of silkworm, Bombyx mori", In Vitro Cell Dev Biol Anim, 45, pp.414~419 (2009)
- 김민성, "Initial Culture Conditions for Primary Cell Populations Derived from Radula Tissue in Abalone Haliotis discus hannai", Fisheries and Aquatic Sciences, 17(3), pp.385~390 (2014), 한국수산과학회
- Kim MS, "Establishment condition and characterization of heart-derived cell culture in Siberian sturgeon(Acipenser baerii)", In Vitro Cell Dev Biol Anim, 50, pp.909~917 (2014)
- Klingseisen A, "Differential and overlapping functions of two closely related Drosophila FGF8-like growth factors in mesoderm development", Development, 136, pp.2393~2402 (2009)
- Lacerda SM, "A new and fast technique to generate offspring after germ cells transplantation in adult fish : the Nile tilapia(Oreochromis niloticus)model. PLoS One 5, e10740", PLoS One, 5, pp.e10740~ (2010)
- Latire T, "Responses of primary cultured haemocytes from the marine gastropod Haliotis tuberculata under 10-day exposure to cadmium chloride", Aquat Toxicol, 109, pp.213~221 (2012)
- Lebel JM, "Effects of different vertebrate growth factors on primary cultures of hemocytes from the gastropod mollusc, Haliotis tuberculata", Biol Cell, 86, pp.67~72 (1996)
- 이동욱, "Establishment and Characterization of Permanent Cell Lines from Oryzias dancena Embryos", Fisheries and Aquatic Sciences, 16(3), pp.177~185 (2013), 한국수산과학회[DOI]
- Naganuma T, "Myogenesis in primary cell cultures from larvae of the abalone, Haliotis rufescens", Mol Mar Biol Biotechnol, 3, pp.131~140 (1994)
- Najmudeen TM, "Gonad maturation of the tropical abalone Haliotis varia Linnaeus 1758(Vetigastropoda : Haliotidae)", Molluscan Res, 27, pp.140~146 (2007)
- Pichon D, "Characterization of abalone Haliotis tuberculata–Vibrio harveyi interactions in gill primary cultures", Cytotechnology, 65, pp.759~772 (2013)
- Poncet JM, "Cryopreservation of mantle dissociated cells from Haliotis tuberculata(Gastropoda)and postthawed primary cell cultures", Cryobiology, 44, pp.38~45 (2002)
- Poncet JM, "In vitro synthesis of proteoglycans and collagen in primary cultures of mantle cells from the nacreous mollusk, Haliotis tuberculata : a new model for study of molluscan extracellular matrix", Mar Biotechnol, 2, pp.387~398 (2000)
- Rhee WJ, "Anti-apoptotic mechanism of silkworm hemolymph in HeLa cell apoptosis", Process Biochem, 48, pp.1375~1380 (2013)
- Sevala VM, "Hemolymph insulin-like peptides(ILP)titers and the influence of ILP and mammalian insulin on the amino acid incorporation in the mantle collar In vitro in Helisoma(Mollusca)", Biol Bull, 185, pp.140~148 (1993)
- Srivatsan M, "Acetylcholinesterase promotes regeneration of neurites in cultured adult neurons of Aplysia", Neuroscience, 77, pp.921~931 (1997)
- Sud D, "Role of water-soluble matrix fraction, extracted from the nacre of Pinctada maxima, in the regulation of cell activity in abalone mantle cell culture(Haliotis tuberculata)", Tiss Cell, 33, pp.154~160 (2001)
- Suja CP, "In vitro culture of mantle tissue of the abalone Haliotis varia Linnaeus", Tiss Cell, 37, pp.1~10 (2005)
- Suja CP, "Effect of culture media and tissue extracts in the mantle explant culture of abalone, Haliotis varia Linnaeus", Aquaculture, 271, pp.516~522 (2007)
- Suwattana D, "In vitro culture of gill and heart tissues of the abalone Haliotis asinina", J Shellfish Res, 29, pp.651~653 (2010)
- van der Merwe M, "Investigating the establishment of primary cell culture from different abalone(Haliotis midae)tissues", Cytotechnology, 62, pp.265~277 (2010)
- Wetakan SA, "Ultrastructure of female germ cells in Haliotis asinina Linnaeus", Invertebr Reprod Dev, 39, pp.67~79 (2001)
- Wong TT, "Production of zebrafish offspring from cultured female germline stem cells", PLoS ONE, 8, pp.e62660~ (2013)
- Yoshino TP, "Molluscan cells in culture : primary cell cultures and cell lines", Can J Zool, 91, pp.391~404 (2013)
- Zou K, "Production of offspring from a germline stem cell line derived from neonatal ovaries", Nat Cell Biol, 11, pp.631~636 (2009)