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Establishment and characterization of a conditionally immortalized smooth muscle/myometrial‐like cell line
Author(s) -
Qian Jin,
Hendrix E. Michael,
Larsen William J.,
Dorn Gerald W.,
Lessard James L.
Publication year - 1997
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199707)47:3<284::aid-mrd7>3.0.co;2-h
Subject(s) - biology , myosin , myometrium , actin , cell culture , microbiology and biotechnology , myocyte , in situ hybridization , antigen , uterus , messenger rna , endocrinology , immunology , gene , biochemistry , genetics
A novel smooth muscle/myometrial‐like cell line, SMU1‐10, has been generated from the uterus of a H‐2K b ‐tsA58 transgenic mouse carrying a thermolabile SV40 large T‐antigen gene. These cells grow continuously when maintained at the permissive temperature (33°C) for the SV40 large T‐antigen but stop dividing when placed at the non‐permissive temperature (39°C) and ultimately die within 3 weeks. All of the SMU1‐10 cells produce smooth muscle α‐actin (SMAA) at both 33°C and 39°C. A subset of the cells also contain smooth muscle γ‐actin (SMGA), a hallmark of smooth muscle differentiation, and the fraction of cells staining for this actin increases from about 1% when maintained for three days at 33°C to as much as 30% at 39°C over the same length of time. However, the appearance of SMGA in SMU1‐10 cells appears to be regulated mainly at a post‐transcriptional level since in situ hybridization indicates that all cells contain SMGA mRNA at both 33°C and 39°C. SMU1‐10 cultures also contain smooth muscle myosin heavy chain (SM‐MHC) and SM22α, both of which are only found in smooth muscle of the adult mouse. Three additional smooth muscle (myometrium)‐related markers, connexin 43, the thromboxane A 2 receptor, and the progesterone receptor also are present in these cells. At the nonpermissive temperature for SV40 large T‐antigen, the both level of SMGA mRNA and the number of cells staining for this actin are significantly increased in the presence of progesterone, a process that is similar to the upregulation of SMGA in the myometrium late in pregnancy. Overall, SMU1‐10 cells provides a potentially useful in vitro model system to study smooth muscle/myometrial differentiation. Mol. Reprod. Dev. 47:284–294, 1997. © 1997 Wiley‐Liss, Inc.