The effect of extracellular matrix molecules on mouse preimplantation embryo development in vitro.
The extracellular matrix (ECM) molecules, laminin (LN), chondroitin sulfate (CS), fibronectin (FN), hyaluronic acid (HA), mucin (MUC) and heparan sulfate proteoglycan (HS), were investigated as supplements to culture medium to improve the in vitro development of mouse 1-cell zygotes to blastocysts. Development was also compared with that in medium supplemented with bovine serum albumin (BSA) to determine the potential for ECM molecules as suitable alternatives to serum albumin in culture medium. Supplementation of sequential culture media with LN at all concentrations examined failed to result in more than 70% of zygotes developing to blastocysts; therefore, LN was considered unsuitable as a replacement for BSA and was not examined further. The optimal concentration of the remaining ECM molecules was used to supplement sequential culture media and the effect on blastocyst quality was assessed by determining the differential cell numbers of blastocysts grown in BSA-supplemented medium. Development to blastocyst was similar, regardless of the macromolecule used. The number of inner cell mass cells was significantly higher in HS-supplemented medium compared with controls. Trophectoderm cell numbers were similar to control values for all ECM molecules examined except CS for which there were fewer trophectoderm cells. It is concluded that ECM molecules, FN, HA, MUC and HS may be used as substitutes for serum protein supplementation of culture media EG0/G2 for mouse preimplantation embryo development. Heparan sulfate proteoglycan increases inner cell mass numbers and this may be due to interactions with the growth factors fibroblast growth factor 4 (FGF-4) and granulocyte-macrophage colony-stimulating factor.
Figueiredo
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F
Monash Institute of Reproduction and Development, Monash University, Monash Medical Centre, 246 Clayton Road, Clayton, Victoria 3168, Australia.
Jones
G M
GM
Thouas
G A
GA
Trounson
A O
AO
D000818:Animals; D001755:Blastocyst / Q000187:drug effects*; D002809:Chondroitin Sulfates / Q000494:pharmacology; D003470:Culture Media; D046508:Culture Techniques; D004305:Dose-Response Relationship, Drug; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D016326:Extracellular Matrix Proteins / Q000494:pharmacology*; D005353:Fibronectins / Q000494:pharmacology; D006025:Glycosaminoglycans / Q000494:pharmacology*; D019812:Heparan Sulfate Proteoglycans / Q000494:pharmacology; D006820:Hyaluronic Acid / Q000494:pharmacology; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009077:Mucins / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D015053:Zygote / Q000187:drug effects*
D003470:Culture Media; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D007797:Laminin; D009077:Mucins; D012710:Serum Albumin, Bovine; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates