DNA methylation patterns of spermatozoa and two generations of offspring obtained after murine spermatogonial stem cell transplantation.
BACKGROUND
Apart from its use in research, spermatogonial stem cell transplantation (SSCT) may have important clinical applications. This controlled study aimed at evaluating the safety of SSCT by analyzing the DNA methylation pattern of Igf2, Peg1 and alpha-Actin both in spermatozoa and live born offspring obtained after SSCT in mice.


METHODS
Testicular cell suspensions were transplanted to the testes of genetically sterile WW recipients. Transplanted males were mated with fertile females and their first and second generation offspring were examined and compared with controls with respect to weight, length and DNA methylation patterns. Sodium-bisulfite treated genomic DNA extracted from post-transplantation spermatozoa, liver, kidney and placenta of first and second generation offspring was PCR-amplified to obtain Igf2, Peg1 and alpha-Actin gene fragments. Pyrosequencing was used to individually quantify the resulting artificial C/T sequence variation at CpG sites.


RESULTS
First and second generation offspring developed normally with their length and weight not being different from controls. Also the DNA methylation patterns of Igf2, Peg1 and alpha-Actin were not different among controls and first and second generation offspring after SSCT.


CONCLUSIONS
SSCT between syngenic individuals was not associated with changes in fetal development nor with differences in the DNA methylation patterns of Igf2, Peg1 and alpha-Actin in spermatozoa or other tissues from two subsequent generations of offspring obtained after SSCT.
Goossens
E
E
Research Laboratory for Embryology and Genetics, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium. ellen.goossens@uzbrussel.be
De Rycke
M
M
Haentjens
P
P
Tournaye
H
H
D000199:Actins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D018899:CpG Islands; D019175:DNA Methylation; D005260:Female; D018392:Genomic Imprinting; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics; D013093:Spermatogonia / Q000378:metabolism*; D013094:Spermatozoa / Q000378:metabolism*; D033581:Stem Cell Transplantation
D000199:Actins; C513121:IGF2 protein, mouse; D011506:Proteins; C103637:mesoderm specific transcript protein; D007335:Insulin-Like Growth Factor II