Response to abiotic and organic substances stimulation belongs to ontologic groups significantly up-regulated in porcine immature oocytes

Abstract

The efficiency of the process of obtaining mature oocytes, and then of porcine embryos in vitro depends on many factors and requires meeting many conditions. These include selection of morphologically appropriate oocytes, selection of appropriate medium components, as well as a number of abiotic factors (appropriate microenvironment during in vitro culture). Oocytes were taken from 45 pubertal crossbred Landrace gilts. The BCB test was carried out. BCB + oocytes were divided into two groups: “before IVM” and “after IVM”. “Before IVM” oocytes were subjected to molecular analyzes immediately after collection, while “after IVM” oocytes underwent in vitro maturation and then the second BCB test. Oocytes that remained BCB+ after the second test were used for molecular analyzes using Affymetrix expression microarrays. A group of genes responsible for response to organic substance and response to abiotic stimulus, which underwent significant changes (decrease) was discovered after oocyte in vitro maturation. Genes such as MM, PLDP, SERPINH, MYOF, DHX9, HSPA5, VCP, KIT, SERPINH1, PLD1, and VCP showed the largest decrease after the culture period. The levels of these genes were therefore elevated in oocytes before the in vitro maturation process. In conclusion, a number of organic and abiotic factors have an impact on the process of the oocyte in vitro maturation. The presented results confirm the literature data in which the low efficiency of obtaining mature oocytes in in vitro conditions is mentioned, which further impacts the amount of viable embryos obtained.

Muscle Cell Morphogenesis, Structure, Development and Differentiation Processes Are Significantly Regulated during Human Ovarian Granulosa Cells In Vitro Cultivation

“…Next, the pellet with GCs was washed twice in culture medium and was centrifugated again with the same settings. The culture medium consisted of DMEM (Sigma; Merck KGaA, Darmstadt, Germany), 2% foetal bovine serum FBS (FBS; Sigma; Merck KGaA, Darmstadt, Germany), 10 mg/mL gentamicin (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 4 mm L-glutamine (stock 200 mm, Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 10,000 μg/mL streptomycin and 10,000 U/mL penicillin (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) [ 15 , 16 , 17 ].…”

Genes Involved in the Processes of Cell Proliferation, Migration, Adhesion, and Tissue Development as New Potential Markers of Porcine Granulosa Cellular ProcessesIn Vitro: A Microarray Approach

“…The Affymetrix procedure was performed as previously described (Kranc et al, 2017(Kranc et al, , 2018a(Kranc et al, , 2018bBryja et al, 2018;Chamier-Gliszczyńska et al, 2018;Stefańska et al, 2018;Nawrocki et al, 2018). cDNA was reverse transcribed from total sample RNA (100 ng) (Ambion Ò WT Expression Kit).…”

Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

“…For amplification, 2 μl of cDNA solution was added to 18 μl of QuantiTect ® SYBR ® Green PCR (Master Mix Qiagen GmbH, Hilden, Germany) and primers ( Table 1). One RNA sample of each preparation was processed without the RT-reaction to provide a negative control for subsequent PCR, where each sample processes in two technical repeats [17][18][19].…”