| 20093282 |
Chen M, Chiou P, Liao YH, Lin CM, Chen T: Development and characterization of five Rainbow Trout pituitary single-cell clone lines capable of producing pituitary hormones. J Endocrinol. 2010 Jan 21.
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 CO2-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 CO2-independent medium supplemented with 10% FBS at 20 degrees C was 3.6 +/- 0.7, 2.8 +/- 0.7, 3.2 +/- 0.8, 5.5 +/- 0.6 and 6.6 +/- 0.6 days, respectively. Each single-cell clone contains 60 +/- 2 chromosomes, which is within the range of the 2N chromosome numbers reported for rainbow trout. Reverse transcription (RT)-PCR analysis revealed that, in addition to expressing growth hormone (GH), prolactin (PRL) and estradiol receptor alpha (E2Ralpha) genes, each single-cell clone line also expressed other pituitary specific genes such as thyroid stimulating hormone (TSH), gonadotropin I (GTH-1), gonadotropin II (GTH-2), somatolactin (SL), proopiomelanocortin-B (POMC-B) and corticoidsteroid receptor (CR). Immunocytochemical analysis showed all five single-cell clones produced both GH and PRL. Furthermore, the expression of GH and PRL genes in the single-cell clone lines are responsive to induction by 17beta-estradiol, dexamethasone and o,p'-dichlorodiphenyltrichloroethane (o,p'-DDT). All together, these results confirm that each of the single-cell 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 also be developed as a rapid screening system for identifying environmental endocrine disruptors. |
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