Long-term culture of Japanese human embryonic stem cells in feeder-free conditions.
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.
Navarro-Alvarez
Nalú
N
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
Soto-Gutierrez
Alejandro
A
Yuasa
Takesui
T
Yamatsuji
Tomoki
T
Shirakawa
Yasuhiro
Y
Nagasaka
Takeshi
T
Sun
Sheng D
SD
Javed
Muhammad Shahid
MS
Tanaka
Noriaki
N
Kobayashi
Naoya
N
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005346:Fibroblast Growth Factors; D018398:Homeodomain Proteins / Q000096:biosynthesis; D006801:Humans; D007564:Japan; D051379:Mice; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D005346:Fibroblast Growth Factors; D000469:Alkaline Phosphatase