Mammalian oviductal epithelial cells have been known to improve in vitro fertilization and embryonic development. Recently, co-cultured human embryos with the epithelial cells in human genital tract has been reported to improve the pregnancy
rate.
The
purpose of the study was to investigate the effects of the epithelial cells of human genital tract on the development of mouse early embryos and human fertilized oocytes.
The epithelial cells of human genital tract were collected from the fallopian tubes which were obtained during hysterectomy in fertile women and from the endometrium during endometrium biopsy. Collected human ampullary cells (HACs) and
endometrial
cells
(HECs) were cultured for 10 days to establish primary monolayer. Second passaged HACs and HECs obtained by trypsinization were cryopreserved in PBS with 1.5M DMSO for later use. To investigate the effect when co-cultured with HACs and HECs, we
tried to
apply strict quality control on mouse embryo, from two cell to blastocyst prior to human trial.
@ES The results of quality control were as follows:
@EN In Group Ⅰ (Ham's F10 with 10% FCS), Group Ⅱ (co-cultured with HACs) and Group Ⅲ (co-cultured with HECs), developmental rates to blastocyst were 63.3% (253/400), 76.0% (304/400), 74/0% (296/400), respectively. Hatching rates were 36.8%
(147/400),
41.8% (167/400), 38/0% (152/400), respectively (p<0.05).
To perform the human IVF, cryopreserved HACs were thawed at 37℃ waterbath, seeded on the well dish and cultured for 48 hrs. The pronuclear stage embryos were transferred to the seeded well dish. After 24 hrs, co-cultured embryos were examined
and
transferred to patient's uterus.
The results of human IVF when co-cultured with HACs were that fertilization and developmental rates were 61.8% (256/414), 95.3% (244256) as compared with 57.2% (279/488) and 94.6% (264/279) in Ham's F10 supplemented with 10% FCS (control).
However,
62.9% (161/256) of co-cultured human embryos showed good embryos (no or slight fragmentation) s compared with 53.8% (150/279) in control (p<0.05). Pregnancy rate was 40.0% (12/30) when co-cultured with HACs whereas 30.6% (11/36) in control.
In conclusions, co-culture system using HACs and HECs improved the developmental and hatching rates of mouse embryo. Also, in human IVF system when co-cultured with HACs, it improved both the quality of human embryos and the pregnancy rate.
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