Alteration and preservation of cellular characteristics in long-term culture of tetraploid H-1 (ES) cells

Fujikawa-Yamamoto, Kohzaburo; Miyagoshi, Minoru; Yamagishi, H; Luo, Xue‐Gang

doi:10.1111/j.1749-0774.2008.00047.x

Cited by 9 publications

(8 citation statements)

References 20 publications

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“…On the contrary, 4H1 cells lost DNA in long-term culturing (Fujikawa-Yamamoto et al, 2008) and octaploid 8H1 cells transformed to hexaploid teratocarcinoma (FujikawaYamamoto et al, 2009). It has been reported that mouse ES cell/fibroblast hybrid cells with a near-tetraploid karyotype yielded diploid/tetraploid chimeras via injection into C57BL mouse blastocysts, suggesting that the DNA of tetraploid hybrid cells was stable following chimera formation (Kruglova et al, 2008).…”

Section: Discussionmentioning

confidence: 96%

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DNA stable pentaploid H1 (ES) cells obtained from an octaploid cell induced from tetraploid cells polyploidized using demecolcine

Fujikawa-Yamamoto

Luo

Miyagoshi

et al. 2010

Journal Cellular Physiology

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Pentaploid H1 (ES) cells (5H1 cells) were accidentally obtained through one-cell cloning of octaploid H1 (ES) cells (8H1 cells) that were established from tetraploid H1 (ES) cells (4H1 cells) polyploidized using demecolcine. The number of chromosomes of 5H1 cells was 100, unlike the 40 of diploid H1 (ES) cells (2H1 cells), 80 of 4H1, and 160 of 8H1 cells. The durations of G(1), S, and G(2)/M phases of 5H1 cells were 3, 7, and 6 h, respectively, almost the same as those of 2H1, 4H1, and 8H1 cells. The cell volume of 5H1 cells was half of that of 8H1 cells, suggesting that 5H1 cells were created through abnormal cell divisions of 8H1 cells. The morphology of growing 5H1 cells was a spherical cluster similar to that of 2H1 cells and differing from the flagstone-like shape of 4H1 and 8H1 cells. Pentaploid solid tumors were formed from 5H1 cells after interperitoneal injection into the mouse abdomen, and they contained endodermal, mesodermal, and ectodermal cells as well as undifferentiated cells, suggesting both that the DNA content of 5H1 cells was retained during tumor formation and that the 5H1 cells were pluripotent. The DNA content of 5H1 cells was stable in long-term culturing as 2H1 cells, meaning that 5H1 and 2H1 cells shared similarities in DNA structure. The excellent stability of the DNA content of 5H1 cells was explained using a hypothesis for the DNA structure of polyploid cells because the pairing of homologous chromosomes in 5H1 cells is spatially forbidden.

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Section: Discussionmentioning

confidence: 96%

“…4H1 cells lost DNA content gradually in long-term culturing (Fujikawa-Yamamoto et al, 2008) or abruptly in DME medium . 8H1 cells showed DNA decay where the octaploid cells changed to hexaploid teratocarcinoma in mouse abdomens 2 months after intraperitoneal injection (Fujikawa-Yamamoto et al, 2009).…”

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confidence: 99%

DNA stable pentaploid H1 (ES) cells obtained from an octaploid cell induced from tetraploid cells polyploidized using demecolcine

Fujikawa-Yamamoto

Luo

Miyagoshi

et al. 2010

Journal Cellular Physiology

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“…4H1 cells lost DNA content gradually in long-term culturing [14] or abruptly in DME medium [15]. 8H1 cells also showed DNA decay where the octaploid cells changed to hexaploid teratocarcinoma [3].…”

Section: Introductionmentioning

confidence: 95%

Hexaploid H1 (ES) cells established from octaploid H1 cells are as DNA stable as pentaploid H1 cells

et al. 2010

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Hexaploid H1 (ES) cells (6H1 cells) were established from octaploid H1 cells (8H1 cells), as were pentaploid H1 cells (5H1 cells). 6H1 cells were compared with 5H1 cells. The number of chromosomes of 6H1 cells was 115, 20 more than the 95 of 5H1 cells. The durations of G(1), S, and G(2)/M phases of 6H1 cells were 3, 7, and 6 h, respectively, almost the same as those of 5H1 cells. The cell volume of 6H1 cells was equivalent that of 5H1 cells. The morphology of 6H1 cells was flattened circular cluster, different from the spherical cluster of 5H1 cells. 6H1 cells exhibited alkaline phosphatase activity as well as 5H1 cells. The DNA content of 6H1 cells was stable and maintained for 300 days of culturing, the same as that of 5H1 cells. The DNA stability of 6H1 cells was explained using a hypothesis concerning the DNA structure of polyploid cells because the asymmetric configuration of homologous chromosomes in 6H1 cells inhibited chromosome loss.

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“…4H1 cells lost DNA gradually in about 180 days [13]. 8H1 cells also showed DNA decay where the octaploid cells changed to hexaploid teratocarcinoma [3].…”

Section: Introductionmentioning

confidence: 96%

Haploid unit-ploidy transition of tetraploid and octaploid H1 (ES) cells in long-term culturing

et al. 2011

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Haploid unit-ploidy transition in tetraploid and octaploid mouse H1 (ES) cells (4H1 and 8H1 cells, respectively) during long-term culturing was observed using flow cytometry. The DNA content of 4H1 cells was elevated from 3.5C to 4.5C, and that of 8H1 cells was degraded from 6.5C to 5.5C, in addition to gradual DNA loss (C: complement). The timing of the transition was not predetermined. Cell cycle parameters, doubling time and phase durations, were essentially the same before and after the transition, suggesting that most cells in a cell population were induced to undergo the ploidy transition at the same time. Cellular morphology was altered before and after the transition, suggesting that the ploidy shift changed cellular characteristics; however, pluripotency was maintained irrespective of DNA content. Cell volume correlated with DNA content during the final stage of culturing. Diploid and hexaploid H1 (ES) cells--2H1 and 6H1 cells, respectively--were used as control cells in which the ploidy was maintained for about 300 days of culturing. The haploid unit-ploidy transition was explained using a hypothesis concerning the DNA structure of polyploid cells: closing homologous chromosomes causes inhomogeneous cell division accompanying a haploid DNA set, suggesting the existence of a coupling apparatus connecting DNA fibers with a single haploid DNA set.

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Alteration and preservation of cellular characteristics in long-term culture of tetraploid H-1 (ES) cells

Cited by 9 publications

References 20 publications

DNA stable pentaploid H1 (ES) cells obtained from an octaploid cell induced from tetraploid cells polyploidized using demecolcine

DNA stable pentaploid H1 (ES) cells obtained from an octaploid cell induced from tetraploid cells polyploidized using demecolcine

Hexaploid H1 (ES) cells established from octaploid H1 cells are as DNA stable as pentaploid H1 cells

Haploid unit-ploidy transition of tetraploid and octaploid H1 (ES) cells in long-term culturing

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