Jurnal Anatomi Indonesia
Aktivitas NADH- tetrazolium reductase sel sel trofoblas pada blastosis yang mengalami hatching dan gagal hatching
Roza Helmita(1*), Ita Djuwita(2), Bambang Purwantara(3), Adi Winarto(4)
(1) Mahasiswa Pascasarjana Institut Pertanian Bogor
(2) BagianAnatomi, Histologi dan Embriologi, DepartemenAnatomi, Fisiologi dan Farmakologi, Fakultas Kedokteran Hewan, Institut Pertanian Bogor
(3) Bagian Fisiologi Reproduksi dan Kebidanan, Departemen Klinik, Reproduksi dan Patologi, Fakultas Kedokteran Hewan, Institut Pertanian Bogor
(4) BagianAnatomi, Histologi dan Embriologi, DepartemenAnatomi, Fisiologi dan Farmakologi, Fakultas Kedokteran Hewan, Institut Pertanian Bogor
(*) Corresponding Author
Abstract
Implantation is themost critical stage in the establishment of pregnancy. Inmammals, it has been estimated that
between 25%and 60%of conceptuses are lost before or at the time of implantation. The objectives of this study
were to investigate the activity of themitochondrial NADH-tetrazoliumreductase, the outgrowth and differentiation
of trophoblast cells in in vitro culture of hatched and non hatched blastocyst. Blastocysts were collected
from mice cornua utery at day-4 of pregnancy and were divided into 3 groups: blastocysts undergo hatching
within 24 hours, 48 hours and non hatching. Embryos were cultured in DMEMmediumin 5%CO2 incubator at
37°C for 10 days.The trophoblastsmonolayer were processed forGimsa staining and histochemistry analysis
of NADH-tetrazolium reductase activity. The outgrowth of trophoblast cells were measured using eyespiece
micrometer. The results showed that the activity of NADH-tetrazolium reductase of 24 h and 48 h hatched
blastocysts showed higher intensity than the non hatched blastocysts (P<0,05). The trophoblast outgrowth
diameter of 24 h hatched blastocystwas significantly higher than the non hatched blastocyst, but not signifcantly
different with the 48 h hatched.Morphology examination using light microscope showed that the trophoblast
monolayer of 24 h hatched blastocyst differentiated into cytotrophoblast, syncytioptrophoblast and
spongiotrophoblast after 5 days of in vitro cultured. In conclusion, in in vitro sudy the failure of blastocysts
implantationwas due to the impairment ofNADHdehydrogenase activity in complex Imitochondria and the failure
of outgrowth and differentiation of the trofoblast cells.
between 25%and 60%of conceptuses are lost before or at the time of implantation. The objectives of this study
were to investigate the activity of themitochondrial NADH-tetrazoliumreductase, the outgrowth and differentiation
of trophoblast cells in in vitro culture of hatched and non hatched blastocyst. Blastocysts were collected
from mice cornua utery at day-4 of pregnancy and were divided into 3 groups: blastocysts undergo hatching
within 24 hours, 48 hours and non hatching. Embryos were cultured in DMEMmediumin 5%CO2 incubator at
37°C for 10 days.The trophoblastsmonolayer were processed forGimsa staining and histochemistry analysis
of NADH-tetrazolium reductase activity. The outgrowth of trophoblast cells were measured using eyespiece
micrometer. The results showed that the activity of NADH-tetrazolium reductase of 24 h and 48 h hatched
blastocysts showed higher intensity than the non hatched blastocysts (P<0,05). The trophoblast outgrowth
diameter of 24 h hatched blastocystwas significantly higher than the non hatched blastocyst, but not signifcantly
different with the 48 h hatched.Morphology examination using light microscope showed that the trophoblast
monolayer of 24 h hatched blastocyst differentiated into cytotrophoblast, syncytioptrophoblast and
spongiotrophoblast after 5 days of in vitro cultured. In conclusion, in in vitro sudy the failure of blastocysts
implantationwas due to the impairment ofNADHdehydrogenase activity in complex Imitochondria and the failure
of outgrowth and differentiation of the trofoblast cells.
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