Chemically fixed nurse cells for culturing murine or primate embryonic stem cells

Ito, Yoshihiro; Kawamorita, Makiko; Yamabe, Toshio; Kiyono, Toru; Miyamoto, Kanji

doi:10.1263/jbb.103.113

Cited by 25 publications

(35 citation statements)

References 23 publications

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“…As we reported, chemically fixed feeder cells supported the growth of hematopoietic stem cells . Therefore, the feeder cells were chemically fixed for supporting the growth of ES cells with keeping the undifferentiated state (Figure 1) [Ito, 2007]. Glutaraldehyde (GA)-or paraformaldehyde (PFA)-fixed MEF cells and HAE cells were prepared for culture of mouse and primate (monkey) ES cells, respectively.…”

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

“…The cells on chemically-fixed cells were stained stronger, even with and without freeze-drying, than that cultured on MEF treated with mitomycine C. On the other hand, the cells cultured only on the gelatin-coated dish were not stained so much. [Ito, 2007] Immunostaining of SSEA-1 and SSEA-4, which are known to be the markers of undifferentiation and differentiation of mouse ES cells, respectively, were performed and the unidifferentiation of mouse ES cells on chemically fixed cells was checked. Expression of transcription factor Oct-3/4 of mouse ES cells was investigated by RT-PCR.…”

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

“…These results confirm the difference between mouse ES cells and primate ES cells in an aspect of self undiferentioation state retaining mechanism. [Ito, 2007] Although fixed HAE cells have incompleteness supporting ability of monkey ES cells, they support the growth and colony formation. Neither chemically fixation nor freeze drying significantly affected the supporting activity, although the growth rates were different from each other.…”

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

“…, and freeze-dried HAE (D) for 3 days, and freeze-dried GA-fixed HAE (E), freeze-dried PFA-fixed HAE (F), and gelatin (G) for 8 days. Reproduced with permission [Ito, 2007] …”

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

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Biomaterials for In Vitro Expansion of Embryonic Stem Cells

Sakuragi¹,

Ito²

2011

Methodological Advances in the Culture, Manipulation and Utilization of Embryonic Stem Cells for Basic and Practical Applicatio

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Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

“…, and freeze-dried HAE (D) for 3 days, and freeze-dried GA-fixed HAE (E), freeze-dried PFA-fixed HAE (F), and gelatin (G) for 8 days. Reproduced with permission [Ito, 2007] …”

Section: Culture Of Es Cells On Chemically Fixed Feeder Cellsmentioning

confidence: 99%

See 2 more Smart Citations

Biomaterials for In Vitro Expansion of Embryonic Stem Cells

Sakuragi¹,

Ito²

2011

Methodological Advances in the Culture, Manipulation and Utilization of Embryonic Stem Cells for Basic and Practical Applicatio

Self Cite

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“…Genbacev et al (2005) isolated human placental fibroblasts from 6-9 weeks of gestation and found that these cells were able to serve as feeder cells to sustain the propagation of human ES cells in an undifferentiated state. Likewise, Miyamoto et al (2004;Ito et al, 2007) cultured human amniotic epithelial cells (hAECs) and human chorionic plate cells derived from placentas and treated them with mitomycin C to mitotically inactivate the cells. It was found that cynomolgus CMK6 cells were able to grow and maintain in an undifferentiated state on these feeder cells.…”

Section: Introductionmentioning

confidence: 99%

Use of Human Amnion Epithelial Cells as a Feeder Layer to Support Undifferentiated Growth of Mouse Embryonic Stem Cells

Lai

Cheng

Liu

et al. 2009

Cloning and Stem Cells

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Mouse and human embryonic stem (ES) cells are usually maintained in an undifferentiated state by coculture with mouse embryonic fibroblasts (MEFs) as feeder cells. In the case of mouse ES cells, addition of leukemia inhibitory factor (LIF) to culture media is also necessary to prevent cell differentiation. Here, we report the use of primary human amnion epithelial cells (hAECs) as feeder cells to culture mouse ES cells using our newly developed protocols. We found that mouse ES cells grown on hAECs express ES cell markers including FGF, Oct-4, Nanog, Sox-2, Rex, and TERT. Interestingly, the expression levels of these genes are three- to five fold higher on hAECs than on MEFs by quantitative real-time PCR. The quicker growing ES cells on hAECs showed a normal 19XY karyotype on passages 25, and ruled out the transformation of ES cells. Using flow cytometry analysis, we show that ES cells grown on hAECs have the same cell cycle distribution pattern as those on MEFs. Further, mouse ES cells cultured on hAECs for at least 20 passages retain the capability of teratoma formation in mice. Finally, we reveal that hAECs express highly LIF that allows for ES growth without the need of addition of commercially obtained LIF. Taken together, our data suggest that hAECs are suitable for mouse ES cell culture and may prove to be a useful alternative to MEFs for human ES cell culture.

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Feeder cells treated with ethanol can be used to maintain self‐renewal and pluripotency of human pluripotent stem cells

et al. 2023

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Feeder cells play an important role in the culture of human pluripotent stem cells (hPSCs) in vitro. Previously, we used methanol as a fixative to prepare feeder cells for the cultivation of pluripotent stem cells (PSCs), and this method could maintain the self‐renewal and pluripotency of PSCs. However, methanol is toxic, and so here we examined whether ethanol could be used to prepare feeder cells as a fixative for hPSC culturing. Primed, naïve, and extended human embryonic stem cells and induced pluripotent stem cells can maintain self‐renewal and undifferentiated potential on feeder cells treated with ethanol for an extended period. RNA sequencing analysis showed that the expression of collagen‐related genes in hPSCs cultured on feeder cells treated with ethanol was significantly lower as compared with hPSCs cultured on feeder cells treated with mitomycin C. Therefore, we speculate that the signaling pathway mediated by collagen‐related genes may, at least in part, contribute to the maintenance of self‐renewal and pluripotency of PSCs induced by feeder cells treated with chemicals.

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Chemically fixed nurse cells for culturing murine or primate embryonic stem cells

Cited by 25 publications

References 23 publications

Biomaterials for In Vitro Expansion of Embryonic Stem Cells

Biomaterials for In Vitro Expansion of Embryonic Stem Cells

Use of Human Amnion Epithelial Cells as a Feeder Layer to Support Undifferentiated Growth of Mouse Embryonic Stem Cells

Feeder cells treated with ethanol can be used to maintain self‐renewal and pluripotency of human pluripotent stem cells

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