Abstract
A major problem in the study of human pituitary cells is their lack of proliferative capacity in vitro. To address this issue, we have infected normal human, postmortem pituitary cells in monolayer culture with a temperature-sensitive (tsA58) mutant of SV40 large T antigen. Several epithelial-like colonies were isolated; and one, designated CHP2, has been studied in detail to identify its functional characteristics. CHP2 cells have undergone more than 150 culture passages and retain an epithelial morphology. They exhibit tight temperature-dependent growth, in the presence and absence of serum, with cell division at 33 C and growth inhibition at 39 C. CHP2 cells, at both temperatures, showed diffuse immunostaining for human α- subunit and focal staining for TSHβ. Gene expression was confirmed by RT- PCR and sequencing. TRH and GnRH receptors Were not detectable, and their absence was confirmed by their lack of effects on intracellular calcium and inositol phospholipids. Cytogenetic analysis showed that the cells had a modal peak in the diploid range and a smaller peak in the tetraploid range. There was also a consistent loss of chromosome 22 and a normal chromosome 2 homologue, the latter being replaced by one of two chromosome 2 markers, M2A or M2B. In conclusion, we have immortalized human pituitary cells using SV40 tsT, from whicH we have cloned a cell line expressing a-subunit and TSHβ.
| Original language | English |
|---|---|
| Pages (from-to) | 1598-1603 |
| Number of pages | 6 |
| Journal | Journal of Clinical Endocrinology and Metabolism |
| Volume | 83 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 1998 |
| Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
- Endocrinology, Diabetes and Metabolism
Cite this
Immortalized human pituitary cells express glycoprotein α-subunit and thyrotropin β (TSHβ). / Ham, J.; Webster, J.; Bond, J. A.; Jasani, B.; Lewis, M. D.; Hepburn, P. J.; Davies, J. S.; Lewis, B. M.; Wynford Thomas, D.; Scanlon, M. F.
In: Journal of Clinical Endocrinology and Metabolism, Vol. 83, No. 5, 1998, p. 1598-1603.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Immortalized human pituitary cells express glycoprotein α-subunit and thyrotropin β (TSHβ)
AU - Ham, J.
AU - Webster, J.
AU - Bond, J. A.
AU - Jasani, B.
AU - Lewis, M. D.
AU - Hepburn, P. J.
AU - Davies, J. S.
AU - Lewis, B. M.
AU - Wynford Thomas, D.
AU - Scanlon, M. F.
PY - 1998
Y1 - 1998
N2 - A major problem in the study of human pituitary cells is their lack of proliferative capacity in vitro. To address this issue, we have infected normal human, postmortem pituitary cells in monolayer culture with a temperature-sensitive (tsA58) mutant of SV40 large T antigen. Several epithelial-like colonies were isolated; and one, designated CHP2, has been studied in detail to identify its functional characteristics. CHP2 cells have undergone more than 150 culture passages and retain an epithelial morphology. They exhibit tight temperature-dependent growth, in the presence and absence of serum, with cell division at 33 C and growth inhibition at 39 C. CHP2 cells, at both temperatures, showed diffuse immunostaining for human α- subunit and focal staining for TSHβ. Gene expression was confirmed by RT- PCR and sequencing. TRH and GnRH receptors Were not detectable, and their absence was confirmed by their lack of effects on intracellular calcium and inositol phospholipids. Cytogenetic analysis showed that the cells had a modal peak in the diploid range and a smaller peak in the tetraploid range. There was also a consistent loss of chromosome 22 and a normal chromosome 2 homologue, the latter being replaced by one of two chromosome 2 markers, M2A or M2B. In conclusion, we have immortalized human pituitary cells using SV40 tsT, from whicH we have cloned a cell line expressing a-subunit and TSHβ.
AB - A major problem in the study of human pituitary cells is their lack of proliferative capacity in vitro. To address this issue, we have infected normal human, postmortem pituitary cells in monolayer culture with a temperature-sensitive (tsA58) mutant of SV40 large T antigen. Several epithelial-like colonies were isolated; and one, designated CHP2, has been studied in detail to identify its functional characteristics. CHP2 cells have undergone more than 150 culture passages and retain an epithelial morphology. They exhibit tight temperature-dependent growth, in the presence and absence of serum, with cell division at 33 C and growth inhibition at 39 C. CHP2 cells, at both temperatures, showed diffuse immunostaining for human α- subunit and focal staining for TSHβ. Gene expression was confirmed by RT- PCR and sequencing. TRH and GnRH receptors Were not detectable, and their absence was confirmed by their lack of effects on intracellular calcium and inositol phospholipids. Cytogenetic analysis showed that the cells had a modal peak in the diploid range and a smaller peak in the tetraploid range. There was also a consistent loss of chromosome 22 and a normal chromosome 2 homologue, the latter being replaced by one of two chromosome 2 markers, M2A or M2B. In conclusion, we have immortalized human pituitary cells using SV40 tsT, from whicH we have cloned a cell line expressing a-subunit and TSHβ.
UR - http://www.scopus.com/inward/record.url?scp=7844246543&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=7844246543&partnerID=8YFLogxK
U2 - 10.1210/jc.83.5.1598
DO - 10.1210/jc.83.5.1598
M3 - Article
VL - 83
SP - 1598
EP - 1603
JO - Journal of Clinical Endocrinology and Metabolism
JF - Journal of Clinical Endocrinology and Metabolism
SN - 0021-972X
IS - 5
ER -