Limits...
Establishment and characterization of immortalized bovine endometrial epithelial cells.

Bai H, Sakurai T, Bai R, Yamakoshi S, Aoki E, Kuse M, Okuda K, Imakawa K - Anim. Sci. J. (2014)

Bottom Line: Immortalized bovine EECs were established by transfecting retroviral vectors encoding human papillomavirus (HPV) E6 and E7, and human telomerase reverse transcriptase (hTERT) genes.In addition, imEECs have maintained the features as EECs, exhibiting oxytocin (OT) and interferon tau (IFNT) responsiveness.Therefore, these imEECs, even after numbers of passages, could be used as an in vitro model to investigate cellular and molecular mechanisms, by which the uterine epithelium responds to IFNT stimulation, the event required for the maternal recognition of pregnancy in the bovine species.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Theriogenology and Animal Breeding, Graduate School of Agricultural and Life Science, the University of Tokyo, Tokyo.

Show MeSH

Related in: MedlinePlus

Proliferation of primary and immortalized endometrial epithelial cells (EECs). Primary (Pri) and immortalized at 60th passage (Im, broken line) EECs were cultured on collagen type IA-coated (+, triangle) or non-coated (−, circle) six-well plates for up to 48 h. The values of CCK-8 (MTT absorbance at 450 nm) were measured at 450 nm at 0, 24, or 48 h (n = 3). Duplicate samples were examined for each treatment, and three independent experiments were performed. Values represent means ± SEM. Statistical significance: **P –; 0.01 vs. Pri (−) within a time period.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4282067&req=5

fig02: Proliferation of primary and immortalized endometrial epithelial cells (EECs). Primary (Pri) and immortalized at 60th passage (Im, broken line) EECs were cultured on collagen type IA-coated (+, triangle) or non-coated (−, circle) six-well plates for up to 48 h. The values of CCK-8 (MTT absorbance at 450 nm) were measured at 450 nm at 0, 24, or 48 h (n = 3). Duplicate samples were examined for each treatment, and three independent experiments were performed. Values represent means ± SEM. Statistical significance: **P –; 0.01 vs. Pri (−) within a time period.

Mentions: ImEECs were grown without apparent signs of senescence for at least 60 passages. Primary EECs at third passages and imEECs at 60th passages were plated onto collagen-coated or non-coated plates and cultured for 24–48 h. Proliferation of primary and immortalized EECs was assessed with the CCK-8 method. As shown in Figure 2, imEECs maintained the proliferation ability equivalent to primary ECCs even after 60 passages (0 h in the figure). Moreover, differences in imEECs' proliferation were seen after 24 h culture and further increase in proliferation was observed at 48 h.


Establishment and characterization of immortalized bovine endometrial epithelial cells.

Bai H, Sakurai T, Bai R, Yamakoshi S, Aoki E, Kuse M, Okuda K, Imakawa K - Anim. Sci. J. (2014)

Proliferation of primary and immortalized endometrial epithelial cells (EECs). Primary (Pri) and immortalized at 60th passage (Im, broken line) EECs were cultured on collagen type IA-coated (+, triangle) or non-coated (−, circle) six-well plates for up to 48 h. The values of CCK-8 (MTT absorbance at 450 nm) were measured at 450 nm at 0, 24, or 48 h (n = 3). Duplicate samples were examined for each treatment, and three independent experiments were performed. Values represent means ± SEM. Statistical significance: **P –; 0.01 vs. Pri (−) within a time period.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4282067&req=5

fig02: Proliferation of primary and immortalized endometrial epithelial cells (EECs). Primary (Pri) and immortalized at 60th passage (Im, broken line) EECs were cultured on collagen type IA-coated (+, triangle) or non-coated (−, circle) six-well plates for up to 48 h. The values of CCK-8 (MTT absorbance at 450 nm) were measured at 450 nm at 0, 24, or 48 h (n = 3). Duplicate samples were examined for each treatment, and three independent experiments were performed. Values represent means ± SEM. Statistical significance: **P –; 0.01 vs. Pri (−) within a time period.
Mentions: ImEECs were grown without apparent signs of senescence for at least 60 passages. Primary EECs at third passages and imEECs at 60th passages were plated onto collagen-coated or non-coated plates and cultured for 24–48 h. Proliferation of primary and immortalized EECs was assessed with the CCK-8 method. As shown in Figure 2, imEECs maintained the proliferation ability equivalent to primary ECCs even after 60 passages (0 h in the figure). Moreover, differences in imEECs' proliferation were seen after 24 h culture and further increase in proliferation was observed at 48 h.

Bottom Line: Immortalized bovine EECs were established by transfecting retroviral vectors encoding human papillomavirus (HPV) E6 and E7, and human telomerase reverse transcriptase (hTERT) genes.In addition, imEECs have maintained the features as EECs, exhibiting oxytocin (OT) and interferon tau (IFNT) responsiveness.Therefore, these imEECs, even after numbers of passages, could be used as an in vitro model to investigate cellular and molecular mechanisms, by which the uterine epithelium responds to IFNT stimulation, the event required for the maternal recognition of pregnancy in the bovine species.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Theriogenology and Animal Breeding, Graduate School of Agricultural and Life Science, the University of Tokyo, Tokyo.

Show MeSH
Related in: MedlinePlus