title: Different responses between diploid and tetraploid H1 embryonic stem cells appeared during long-term culturing in L15F10 medium without leukemia inhibitory factor.
abstract: To examine the alteration in cellular characteristics of polyploid embryonic stem (ES) cells during long-term culturing without leukemia inhibitory factor (LIF), mouse diploid and tetraploid H-1 (ES) cells (2H1 and 4H1 cells, respectively) were cultured without LIF for approximately 5 months. 2H1 and 4H1 cells were adapted to the medium without LIF by decreasing the concentration for several passages, and they were denoted as 2H1(⁻) and 4H1(⁻) cells, respectively. DNA content of 4H1(⁻) cells decreased gradually in the early stage, increased abruptly in the second stage, and then was maintained for a long time. 4H1(⁻) cells exhibited longer doubling time and equivalent phase fraction compared with those of 2H1(⁻) cells. The G₁ phase fractions of 2H1(⁻) and 4H1(⁻) cells were increased compared with that of 2H1 cells. Cellular morphology and pluripotency were maintained in 4H1(⁻) cells but not in 2H1(⁻) cells. 2H1(⁻) cells showed a cell population consisting of several kinds of cells, and they lost alkaline phosphatase activity, suggesting that the cells had differentiated. 4H1(⁻) cells, however, exhibited alkaline phosphatase activity and formed teratocarcinoma in mouse abdomen, suggesting that the cells maintained their pluripotency in the medium without LIF.
authors.affiliation: Division of Cell Medicine, Research Institute of Medical Science, Kanazawa Medical University, Uchinada, Ishikawa, Japan.
authors.forename: Xianwen
authors.initials: X
authors.lastname: Luo
authors.forename: Kohzaburo
authors.initials: K
authors.lastname: Fujikawa-Yamamoto
authors.forename: Minoru
authors.initials: M
authors.lastname: Miyagoshi
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yamagishi
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002454:Cell Differentiation; D002455:Cell Division; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D003470:Culture Media; D004171:Diploidy; D004622:Embryo, Mammalian / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000201:enzymology / Q000473:pathology; D053605:Leukemia Inhibitory Factor; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009368:Neoplasm Transplantation; D018243:Teratocarcinoma / Q000473:pathology; D057891:Tetraploidy; D013997:Time Factors
chemical_list: D003470:Culture Media; D053605:Leukemia Inhibitory Factor; D000469:Alkaline Phosphatase