THE INFLUENCE OF 3D CULTURE CELL ON THE MATURATION OF PREIMPLANTATION MOUSE EMBRYOS IN VITRO
Journal Title: Naukovyi visnyk Chernivetskoho universytetu. Biolohiia (Biolohichni systemy) - Year 2017, Vol 9, Issue 1
Abstract
Creating of conditions for the growth of cells on two-dimensional surfaces, that would match their natural threedimensional conditions in the body tissues, is possible using surfaces modified by different polymers. The cultivation of cells on such nanosurfaces provides optimal conditions closest to conditions in vivo and determines all further cell growth processes, their differentiation, migration and synthesis of the components in intercellular matrix. The research aim was to investigate the effect of modified nanosurfaces, grafted by albumin and biogel, on the morphokinetic characteristics of mouse 2-cell embryos cultivated in vitro. For experiments, mouse pre-implantation embryos were removed from еру mice females after superovulation with intra-peritoneal injection (i.p.) of 5 IU pregnant mare’s serum gonadotropin (PMSG; Sigma Chemical, MO, USA) and 5 IU of human chorionic gonadotropin (hCG; Pregnyl, Organon, Netherlands) in the 48 h interval and were housed in male mice. The 2-cells embryos were transferred into 500 μldrops of KSMO medium supplemented with 5 % FCS and Oviduct Epithelial Cells Culture (OECs) (0.5х106 cells/ml) (control group), group 1 – biogel+KSMO+MOECs; group 2 – albumin+KSMO+ MOECs. The efficiency of three-dimensional oviduct cell culture on biogel and albumin was determined by the development of pre-implantation embryos to the blastocyst stage in vitro and biochemical indexes of the condition medium. During co-culturing on the modified nanosurfaces by biogel or albumin with the layer of Oviduct Epithelial Cells Culture (OECs), the 4-6-cell stage at 24 hours was reached by respectively 26.7 and 20% more embryos versus those for the control group. To 48 hours of culturing the morula stage was reached by respectively 33.3% and 26.7% embryos in control and experimental group with albumin versus 53.3% those embryos in experimental group with biogel. On the third day in experimental groups during culturing onto both nanosurfaces with cells monolayer 6.7% and 13.3% of the embryos were compacted unlike in the control group. Comparison of the results of two-cell mice embryos development in the control and experimental groups showed that cultivation on nanosurfaces with albumin, and especially with biogel, promotes better embryos development.
Authors and Affiliations
О. В. ШТАПЕНКО, Н. М. МАТВІЄНКО
Keywords
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