Long-term culture of Japanese human embryonic stem cells in feeder-free conditions

Nalú Navarro-Alvarez, Alejandro Soto-Gutierrez, Takesui Yuasa, Tomoki Yamatsuji, Yasuhiro Shirakawa, Takeshi Nagasaka, Sheng Sun D, Muhammad Shahid Javed, Noriaki Tanaka, Naoya Kobayashi

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Human pluripotent embryonic stem cells (hESCs) have great promise for research into human developmental biology, development of cell therapies for the treatment of diseases, toxicology, and drug discovery. Traditionally, undifferentiated hESCs are maintained on mouse embryonic fibroblasts (MEFs), which impede the clinical applications of hESCs. Here we have examined the long-term stability of the Japanese hESC line (KhES-1) in feeder-free culture. KhES-1 cells were cultured with MEF conditioned medium (CM) and different doses of basic fibroblast growth factor (bFGF) in six-well-plates of which the surface was coated with Matrigel. KhES-1 cells were maintained for at least 40 passages. In this culture system, the cells maintained stable proliferation rates and steadily expressed Oct-4, Nanog, and alkaline phosphatase. In addition, KhES-1 cells maintained without direct feeder contact formed embryonic bodies with expression of markers from the three germ layers. Here we demonstrated that Japanese human embryonic stem cells KhES-1 were cultured long term in a feeder-free method, while retaining pluripotency in vitro.

Original languageEnglish
Pages (from-to)27-33
Number of pages7
JournalCell Transplantation
Volume17
Issue number1-2
DOIs
Publication statusPublished - Jan 1 2008

Keywords

  • Basic fibroblast growth factor (bFGF)
  • Human ES cells
  • Undifferentiation

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

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  • Cite this

    Navarro-Alvarez, N., Soto-Gutierrez, A., Yuasa, T., Yamatsuji, T., Shirakawa, Y., Nagasaka, T., Sun D, S., Javed, M. S., Tanaka, N., & Kobayashi, N. (2008). Long-term culture of Japanese human embryonic stem cells in feeder-free conditions. Cell Transplantation, 17(1-2), 27-33. https://doi.org/10.3727/000000008783906900