Fish cell culture: Initiation of a line of pituitary cells from carp (Cyprinus carpio) to study the release of gonadotropin in vitro

Ribeiro, Luiz P.; Ahne, W.

doi:10.1007/bf02796467

Cited by 16 publications

(3 citation statements)

References 9 publications

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“…Recently pituitary fragments in a perifusion system have been described in goldfish (Mackenzie et al, 1984). The use df dispersed pituitary cells of the whole pituitary has been also reported either in short-term incubation (rainbow trout: Fihraeus-van Ree et al, 1982a,b) or in primary culture in a static model (rainbow trout: Fkhraeusvan Ree et al, 1982b; carp: Ribeiro and Ahne, 1982;Ribeiro et al, 1983) and in a perifusion system (goldfish: Chang et al, 1984). Recently a primary culture of purified pituitary gonadotrophs have been set up for African catfish (Leeuw et al, 1984).…”

Section: Dlscusslonmentioning

confidence: 99%

Use of pituitary cells in primary culture to study the regulation of gonadotropin hormone (GtH) secretion in rainbow trout: Setting up and validating the system as assessed by its responsiveness to mammalian and salmon gonadotropin releasing hormone

Weil

Hansen

Hyam

et al. 1986

General and Comparative Endocrinology

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To study the regulation of gonadotropin secretion in rainbow trout in vitro, a method for preparing primary cultures of dispersed pituitary cells is described. Cells were dispersed by collagenase 0.1% in Hank's saline solution for 20 hr at 12 degrees and a high yield of viable cells was obtained. Attempts to improve cell functioning were made by varying culture conditions (density of cells initially plated, age of the culture). Cell functioning was assessed by their ability to respond to increasing doses of mammalian and salmon GnRH. Pituitaries were collected from spermiating males whose pituitaries are known to be sensitive to mammalian GnRH in vivo. Using 96-well plates, optimal conditions for good biological activity, are initial plating with 6.2 X 10(4) cells, incubation with GnRH for 24 hr on the third day after plating. In these conditions mammalian analog and salmon GnRH induced an increase in GtH release for doses ranging from 10(-9) to 10(-6) M. The GtH released during the GnRH incubation period does not decrease the sensitivity of the system since addition of 20 ng of GtH at the beginning of incubation does not modify the response profile.

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Section: Dlscusslonmentioning

confidence: 99%

Use of pituitary cells in primary culture to study the regulation of gonadotropin hormone (GtH) secretion in rainbow trout: Setting up and validating the system as assessed by its responsiveness to mammalian and salmon gonadotropin releasing hormone

Weil

Hansen

Hyam

et al. 1986

General and Comparative Endocrinology

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“…Almost all these problems could be overcome if continuous pituitary cell lines that can produce pituitary peptide hormones are available. Unfortunately, very few fish pituitary cell lines are available to date (Ribeiro & Ahne 1982, Bols & Lee 1991, Bols et al 1995.…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of five rainbow trout pituitary single-cell clone lines capable of producing pituitary hormones

Chen¹,

Chiou²,

Liao³

et al. 2010

Journal of Endocrinology

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Five single-cell clone lines (mRTP1B, mRTP1E, mRTP1F, mRTP1K, and mRTP2A) have been developed from adult rainbow trout pituitary glands. These cell lines have been maintained in a CO 2 -independent medium supplemented with 10% fetal bovine serum (FBS) for more than 150 passages. At about 150 passages, the doubling time of each single-cell clone in a CO 2 -independent medium supplemented with 10% FBS at 20 8C was 3 . 6G0 . 7, 2 . 8G0 . 7, 3 . 2G0 . 8, 5 . 5G0 . 6, and 6 . 6G0 . 6 days respectively. Each single-cell clone contains 60G2 chromosomes, which is within the range of the 2N chromosome numbers reported for rainbow trout. Reverse transcription-PCR analysis revealed that in addition to expressing gh, prolactin (prl ), and estradiol (E 2 ) receptor a (e2ra or esr1) genes, each single-cell clone line also expressed other pituitary-specific genes such as tsh, gonadotropin 1 (gth-1 or fshb), gonadotropin 2 (gth-2 or lhb), somatolactin (sl or smtl ), proopiomelanocortin-B (pomcb), and corticosteroid receptor (cr or nr3c1). Immunocytochemical analysis showed that all the five single-cell clones produced both Gh and Prl. Furthermore, the expression of gh and prl genes in the single-cell clone lines is responsive to induction by E 2 , dexamethasone, and o,p 0 -dichlorodiphenyltrichloroethane. All together, these results confirm that each of the singlecell clones was derived from rainbow trout pituitary glands. These single-cell clone lines not only can be used to study factors that regulate the expression of pituitary hormone genes, but can also be developed as a rapid screening system for identifying environmental endocrine disruptors.

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“…In der vorliegenden Arbeit wurde versucht, aus Karpfenhypophysenzellen, die prirnar der Untersuchung der Gonadotropinfreisetzung in vitro dienten (16), einen fur die Fischvirologie geeigneten Zellstamm (CaPi) zu etablieren.…”

Section: Introductionunclassified

Etablierung eines fischvirus empfänglichen zellstammes aus Hypophysen des Karpfens (Cyprinus carpio L.)

Ribeiro

Ahne

1983

Zentralblatt für Bakteriologie, Mikrobiologie und Hygiene. 1. A

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A new continous cell strain (CaPi) was established from normal pitu itary tissue of carp, Cyprinus carpio L.Cells of a frac tio na ted dispersion (0.25% tryp sin-PBS, 10 "C) of pit uitar ies for med mon olayer (fibroblast-and epithelial-lik e-cells) at 25°C using Eagle's mini mal essentia l medium (M EM) supp lemented with 10% fetal calf serum and non essential amino acids . The continous cell strai n, designated CaPi, was obtained by an enzymati cal selecti on of epitheloid cells fr om the monolayer of the primary pit uitary cell cult ure. T his culture has been subculter ed 70 times over a period of 24 mon ths . CaPi-c ells multiply over a temperature range of 15-35°C wi th an optimum growth temperature of 30°C at a seedi ng density of 1.7 x 10· cells /m\. Chromosome ana lysis ind icate s th e cells are het eroploid and show modal numb ers of 47 chromoso mes. T he CaPi cell line is susceptible to fishviru ses (VHSV, PFR, SVCV) producing cyto pathic effects (CPE). H owever, IPNV repl icate s witho ut visible cell alterations.

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Fish cell culture: Initiation of a line of pituitary cells from carp (Cyprinus carpio) to study the release of gonadotropin in vitro

Cited by 16 publications

References 9 publications

Use of pituitary cells in primary culture to study the regulation of gonadotropin hormone (GtH) secretion in rainbow trout: Setting up and validating the system as assessed by its responsiveness to mammalian and salmon gonadotropin releasing hormone

Use of pituitary cells in primary culture to study the regulation of gonadotropin hormone (GtH) secretion in rainbow trout: Setting up and validating the system as assessed by its responsiveness to mammalian and salmon gonadotropin releasing hormone

Development and characterization of five rainbow trout pituitary single-cell clone lines capable of producing pituitary hormones

Etablierung eines fischvirus empfänglichen zellstammes aus Hypophysen des Karpfens (Cyprinus carpio L.)

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