Derivation of human embryonic stem cell lines from embryos obtained after IVF and after PGD for monogenic disorders.
BACKGROUND
Human embryonic stem (hES) cells are pluripotent cells usually derived from the inner cell mass (ICM) of blastocysts. Because of their ability to differentiate into all three embryonic germ layers, hES cells represent an important material for studying developmental biology and cell replacement therapy. hES cell lines derived from blastocysts diagnosed as carrying a genetic disorder after PGD represent in vitro disease models.


METHODS
ICMs isolated by immunosurgery from human blastocysts donated for research after IVF cycles and after PGD were plated in serum-free medium (except VUB01) on mouse feeder layers.


RESULTS
Five hES cell lines were isolated, two from IVF embryos and three from PGD embryos. All lines behave similarly in culture and present a normal karyotype. The lines express all the markers considered characteristic of undifferentiated hES cells and were proven to be pluripotent both in vitro and in vivo (ongoing for VUB05_HD).


CONCLUSIONS
We report here on the derivation of two hES cell lines presumed to be genetically normal (VUB01 and VUB02) and three hES cell lines carrying mutations for myotonic dystrophy type 1 (VUB03_DM1), cystic fibrosis (VUB04_CF) and Huntington disease (VUB05_HD).
Mateizel
I
I
Research Centre for Reproduction and Genetics, University Hospital and Medical School of the Vrije Universiteit Brussel (Dutch-speaking Brussels Free University) Laarbeeklaan 101, 1090 Brussels, Belgium.
De Temmerman
N
N
Ullmann
U
U
Cauffman
G
G
Sermon
K
K
Van de Velde
H
H
De Rycke
M
M
Degreef
E
E
Devroey
P
P
Liebaers
I
I
Van Steirteghem
A
A
D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D003550:Cystic Fibrosis / Q000175:diagnosis / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005307:Fertilization in Vitro; D030342:Genetic Diseases, Inborn / Q000175:diagnosis*; D006801:Humans; D006816:Huntington Disease / Q000175:diagnosis / Q000235:genetics; D009223:Myotonic Dystrophy / Q000175:diagnosis / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology*; D011247:Pregnancy; D019836:Preimplantation Diagnosis
D015415:Biomarkers