title: Effects of group culture on the development of discarded human embryos and the construction of human embryonic stem cell lines.
abstract: OBJECTIVE
  To explore the effect of group culture on the developmental potential of discarded embryos in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles and establish the human embryonic stem cell lines for future research.
  
  
  METHODS
  Fresh discarded embryos were collected from the IVF/ICSI-ET program in the reproductive medical center of the first affiliated hospital of Zhengzhou university in this study. All zygotes were individually cultured from Day 1 to Day 3. On Day 3, discard embryos were then cultured in group of 1-4 embryos per droplet (30 μl/droplets) with a constant culture medium until Day 5 or 6. Mechanical method was used to isolate the inner cell mass (ICM) of blastocyst from the embryo. Then we inoculated the ICM on feeder layer. After identification of those cells, the human embryonic stem cell lines (hESCs) were established.
  
  
  RESULTS
  In this study, we collected 1,223 fresh discarded embryos and they were sequential cultured to the blastocysts (18.07 %, 221/1,223), in which good quality blastocysts were 61(4.98 %, 61/1,223). There was no significant difference in the patients. The embryos from 1PN, 2PN, 3PN were sequential cultured to the blastocyst s(39.31 %,92/234;12.87 %,64/497;13.21 %,65/492),in which good quality blastocysts was 13.6 %(32/92),2.61 %(13/64), 3.04 %(15/65).1PN embryo's blastulation rate and quality embryo formation rate was significantly higher than the 2PN and 3PN embryos' (P <0.05). Three embryos group cultivation has the highest blastulation rate and quality embryo formation rate (P <0.05). In total, we successfully established 4 hESCs lines.
  
  
  CONCLUSIONS
  The group culture of human discard embryos can improve the blastulation rate and blastocyst quality to some extent. Three embryos group cultivate is the better culture number. Human discard embryos are good source for establishment of hESCs.
authors.affiliation: Reproductive Medical Center, First Affiliated Hospital of Zhengzhou University, Constructive East Road, Zhengzhou, 450052, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Sun
authors.forename: Wenzhu
authors.initials: W
authors.lastname: Yu
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Wenyan
authors.initials: W
authors.lastname: Song
authors.forename: Haixia
authors.initials: H
authors.lastname: Jin
authors.forename: Yingpu
authors.initials: Y
authors.lastname: Sun
mesh_terms: D000328:Adult; D001755:Blastocyst / Q000502:physiology*; D002460:Cell Line; D003470:Culture Media / Q000378:metabolism; D046149:Embryo Culture Techniques / Q000379:methods*; D047108:Embryonic Development / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000502:physiology*; D061252:Feeder Cells / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D006801:Humans; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods; D055815:Young Adult; D015053:Zygote / Q000502:physiology
chemical_list: D003470:Culture Media
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