title: Extended Culture of Encapsulated Human Blastocysts in Alginate Hydrogel Containing Decidualized Endometrial Stromal Cells in the Presence of Melatonin.
abstract: Extended in vitro culture of human embryos beyond blastocyst stage could serve as a tool to explore the molecular and physiological mechanisms underlying embryo development and to identify factors regulating pregnancy outcomes. This study presents the first report on the maintenance of human embryo in vitro by alginate co-encapsulation of human blastocyst and decidualized endometrial stromal cells (EnSCs) under melatonin-fortified culture conditions. The effectiveness of the 3D culture system was studied through monitoring of embryo development in terms of survival time, viability, morphological changes, and production of the two hormones of 17b-oestradiol and human chorionic gonadotropin. The embryo structural integrity was preserved during alginate encapsulation; however, only 23 % of the encapsulated embryos could retain in the hydrogels over time and survived until day 4 post-encapsulation. The culture medium fortification with melatonin significantly elevated the maintenance rate of expanded embryos in alginate beads by 65 % and prolonged survival time of human embryos to day 5. Furthermore, embryo co-culture with EnSCs using melatonin-fortified medium increased the survival time of encapsulated embryos to 44 %. The levels of two measured hormones significantly rose at day 4 in comparison with day 2 post-encapsulation especially in the group co-encapsulated with EnSCs and cultivated in melatonin-fortified culture medium. These data are the first evidence representing in vitro development of human embryos until day 10 post-fertilization. This achievement can facilitate the investigation of the mechanisms regulating human embryo development.
authors: Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, P.O. Box: 1177-19615, Tehran, Iran.
authors: Fatemeh
authors: F
authors: Arjmand
authors: Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, P.O. Box: 1177-19615, Tehran, Iran.
authors: Manijeh
authors: M
authors: Khanmohammadi
authors: Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, P.O. Box: 1177-19615, Tehran, Iran.
authors: Shaghayegh
authors: S
authors: Arasteh
authors: Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, P.O. Box: 1177-19615, Tehran, Iran.
authors: Afsaneh
authors: A
authors: Mohammadzadeh
authors: Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, P.O. Box: 1177-19615, Tehran, Iran. s.kazemnejad@ari.ir.
authors: Somaieh
authors: S
authors: Kazemnejad
authors: Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, P.O. Box: 1177-19615, Tehran, Iran. akhondi@ari.ir.
authors: Mohammad-Mehdi
authors: MM
authors: Akhondi
mesh_terms: D000328:Adult; D000464:Alginates / Q000737:chemistry*; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects; D018920:Coculture Techniques; D003470:Culture Media / Q000737:chemistry; D046149:Embryo Culture Techniques / Q000379:methods*; D047108:Embryonic Development; D004717:Endometrium / Q000166:cytology*; D005260:Female; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D008550:Melatonin / Q000494:pharmacology*; D011247:Pregnancy; D017154:Stromal Cells / Q000166:cytology*
chemical_list: D000464:Alginates; D003470:Culture Media; D006603:Hexuronic Acids; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid; D008550:Melatonin
keywords: Alginate hydrogel; Co-culture; Endometrial stromal cell; Human blastocyst; Melatonin