A METHOD FOR OBTAINING A XENO-FREE HBS CELL LINE

A method for obtaining a stable xeno-free hBS cell line, xeno-free hBS cell lines obtained according to said method and use thereof. The method comprises the steps of: i) removing the zona pellucida from a blastocyst to obtain trophectoderm-enclosed inner cell mass by a xeno-free procedure, ii) at l...

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Hauptverfasser: STREHL, RAIMUND, HYLLNER, SVEN JOHAN, KILMARE, EVA KARIN, SEMB, HENRIK, MOYA, KARINA, ELLERSTROEM, CATHARINA, FREJ, KATARINA
Format: Patent
Sprache:eng ; fre ; ger
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Zusammenfassung:A method for obtaining a stable xeno-free hBS cell line, xeno-free hBS cell lines obtained according to said method and use thereof. The method comprises the steps of: i) removing the zona pellucida from a blastocyst to obtain trophectoderm-enclosed inner cell mass by a xeno-free procedure, ii) at least partly removing the trophectoderm to obtain isolated inner cell mass cells by a xeno-free procedure, iii) placing the inner cell mass cells on a layer of human feeder cells in a xeno-free medium, iv) co-culturing of the inner cell mass cells with human feeder cells for a time period of from about 5 days to about 50 days in a xeno-free medium, v) releasing the inner cell mass cells or cells derived thereof from trophectoderm overgrowth, if any, by a xeno-free procedure, vi) selectively, transferring the inner cell mass cells or cells derived thereof to a fresh layer of human feeder cells in a xeno-free medium to obtain xeno-free hBS cells, vii) propagating the xeno-free hBS cells by co-culturing with human feeder cells in a xeno-free medium to obtain a xeno-free hBS cell line. The xeno-free hBS cell line is suitable for use in medicine and in in vitro testing.