Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Development of pre-implantation porcine blastocysts cultured within alginate hydrogel systems either supplemented with secreted phosphoprotein 1 or conjugated with Arg-Gly-Asp Peptide

Taylor D. Laughlin A , Jeremy R. Miles B C , Elane C. Wright-Johnson B , Lea A. Rempel B , Clay A. Lents B and Angela K. Pannier A C
+ Author Affiliations
- Author Affiliations

A Department of Biological Systems Engineering, University of Nebraska – Lincoln, PO Box 830726, Lincoln, NE 68583, USA.

B USDA1, U.S. Meat Animal Research Center (USMARC), PO Box 166, Clay Center, Nebraska 68933, USA.

C Corresponding authors. Email: jeremy.miles@ars.usda.gov; apannier2@unl.edu

Reproduction, Fertility and Development - https://doi.org/10.1071/RD16366
Submitted: 15 September 2016  Accepted: 14 March 2017   Published online: 28 April 2017

Abstract

Although deficiencies in porcine blastocyst elongation play a significant role in early embryonic mortality and establishment of within-litter developmental variation, the exact mechanisms of elongation are poorly understood. Secreted phosphoprotein 1 (SPP1) is increased within the uterine milieu during early porcine pregnancy and contains an Arg-Gly-Asp (RGD) peptide sequence that binds to cell surface integrins on the uterine endometrium and trophectoderm, promoting cell adhesion and migration. The aim of the present study was to evaluate the development of preimplantation porcine blastocysts encapsulated and cultured within alginate hydrogels either supplemented with SPP1 or conjugated with RGD. Blastocysts encapsulated within alginate hydrogels supplemented with SPP1 or conjugated with RGD had increased survival compared with non-encapsulated control blastocysts. In addition, the percentage of blastocysts encapsulated within RGD hydrogels that underwent morphological changes was greater than that of blastocysts encapsulated within standard alginate hydrogels or SPP1-supplemented hydrogels. Finally, only blastocysts encapsulated within RGD hydrogels had both increased expression of steroidogenic and immune responsiveness transcripts and increased 17β-oestradiol production, consistent with blastocysts undergoing elongation in vivo. These results illustrate the importance of the integrin-binding RGD peptide sequence for stimulating the initiation of blastocyst elongation.

Additional keywords: embryo elongation, embryology, gene expression, oestrogen.


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