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Large-scale propagation of four undifferentiated human embryonic stem cell lines in a feeder-free culture system.

Författare och institution:
Eva Sjögren-Jansson (Institutionen för laboratoriemedicin, Avdelningen för klinisk kemi/transfusionsmedicin); Mathilda Zetterström (Institutionen för klinisk neurovetenskap); Karina Moya (-); Jenny Lindqvist (-); Raimund Strehl (-); Peter S Eriksson (Institutionen för klinisk neurovetenskap)
Publicerad i:
Developmental dynamics : an official publication of the American Association of Anatomists, 233 ( 4 ) s. 1304-14
Artikel, refereegranskad vetenskaplig
Sammanfattning (abstract):
We describe an improved and more robust protocol for transfer and subsequent propagation of human embryonic stem cells under feeder-free conditions. The results show that mechanical dissociation for transfer of the human embryonic stem cells to Matrigel resulted in highest survival rates. For passage of the cultures on the other hand, enzymatic dissociation was found to be most efficient. In addition, this method reduces the time, work, and skills needed for propagation of the human embryonic stem cells. With the present protocol, the human embryonic stem cells have been cultured under feeder-free conditions for up to 35 passages while maintaining a normal karyotype, stable proliferation rate, and high telomerase activity. Furthermore, the feeder-free human embryonic stem cell cultures express the transcription factor Oct-4, alkaline phosphatase, and cell surface markers SSEA-3, SSEA-4, Tra 1-60, Tra 1-81, and formed teratomas in severe combined immunodeficient mice. This method provides distinct advantages compared with previous protocols and make propagation of human embryonic stem cells less laborious and more efficient.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
Cell Culture Techniques, Cell Differentiation, genetics, Cell Line, Collagen, Drug Combinations, Embryo, Mammalian, cytology, enzymology, Humans, Immunohistochemistry, In Situ Hybridization, Fluorescence, Karyotyping, Laminin, Proteoglycans, Reverse Transcriptase Polymerase Chain Reaction, Stem Cells, enzymology, Telomerase, metabolism, Teratoma, Tumor Cells, Cultured
Postens nummer:
Posten skapad:
2009-09-09 14:20
Posten ändrad:
2010-03-05 14:25

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