title: Increasing of blastocyst rate and gene expression in co-culture of bovine embryos with adult adipose tissue-derived mesenchymal stem cells.
abstract: OBJECTIVE
  Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development.
  
  
  METHODS
  In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 103and 104cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 104b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran).
  
  
  RESULTS
  In experiment 1, co-culture with 104b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 103b-ATMSCs/mL (p = 0.051), group 104b-ATMSCs/mL yielded higher results of blastocyst production. In experiment 2, when compared to group Gran, co-culture with 104b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05).
  
  
  CONCLUSIONS
  Co-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in bovine and other species.
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil. moyses@ufpa.br.|Laboratório de Células Tronco em Medicina Veterinária (LabTron-Vet), Instituto de Medicina Veterinária, Universidade Federal do Pará, Castanhal, PA, Brazil. moyses@ufpa.br.
authors.forename: Moysés S
authors.initials: MS
authors.lastname: Miranda
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil.
authors.forename: Hamilton S
authors.initials: HS
authors.lastname: Nascimento
authors.affiliation: Laboratório de Células Tronco em Medicina Veterinária (LabTron-Vet), Instituto de Medicina Veterinária, Universidade Federal do Pará, Castanhal, PA, Brazil.
authors.forename: Mayra P R
authors.initials: MP
authors.lastname: Costa
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil.
authors.forename: Nathália N
authors.initials: NN
authors.lastname: Costa
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil.
authors.forename: Karynne N L
authors.initials: KN
authors.lastname: Brito
authors.affiliation: Laboratório de Células Tronco em Medicina Veterinária (LabTron-Vet), Instituto de Medicina Veterinária, Universidade Federal do Pará, Castanhal, PA, Brazil.
authors.forename: Cinthia T A
authors.initials: CT
authors.lastname: Lopes
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil.
authors.forename: Simone S D
authors.initials: SS
authors.lastname: Santos
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil.
authors.forename: Marcela S
authors.initials: MS
authors.lastname: Cordeiro
authors.affiliation: Laboratório de Fecundação In Vitro, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil.
authors.forename: Otávio M
authors.initials: OM
authors.lastname: Ohashi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000328:Adult; D000818:Animals; D001755:Blastocyst / Q000187:drug effects; D002417:Cattle; D018920:Coculture Techniques; D003470:Culture Media; D047108:Embryonic Development; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006107:Granulosa Cells / Q000166:cytology; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D011247:Pregnancy
chemical_list: D003470:Culture Media; D009004:Monosaccharide Transport Proteins; D050814:Octamer Transcription Factor-3
keywords: Bovine; Co-culture; Embryo; IVEP; Mesenchymal stem cells