Influence of seasonal changes on the karyotype stability of Ukrainian Black and White dairy cows

Of the total dairy cattle, 49 % belong to the Ukrainian black-and-white dairy breed. Animals are demanding to the conditions of detention, so the aim of the study was to study the impact of seasonal changes in the environment on the stability of the karyotype of cows of three age groups (first-born, adult and cows 8–10 years). The variability of the karyotype of the studied animals in different seasons of the year was manifested in the form of quantitative and structural disorders of chromosomes. Of the total dairy cattle, 49 % belong to the Ukrainian black-and-white dairy breed. Animals are demanding to the conditions of detention, so the aim of the study was to study the impact of seasonal changes in the environment on the stability of the karyotype of cows of three age groups (first-born, adult and cows 8–10 years). The variability of the karyotype of the studied animals in different seasons of the year was manifested in the form of quantitative and structural disorders of chromosomes. In primiparous women, the most pronounced genomic variability (aneuploidy) was observed in summer and winter and was 16.4 % and 8.8 %. In summer, this variability was more than twice the limit of spontaneous chromosomal variability. The highest percentage of structural chromosome abnormalities (chromosomal breaks) in cows of three age groups was observed in winter and was 2.5 %, 2.8 %, 3.0 %, respectively, and did not exceed the limit of spontaneous mutagenesis characteristic of cattle. An increased proportion of lymphocytes with a micronucleus in first-borns and cows aged 8–10 years (5.6 ‰, 6.4 ‰, respectively) was observed in the summer. The increased proportion of dinuclear lymphocytes appeared in the first-born in the summer season and amounted to 7.6 ‰ (at P > 0.99), and in cows 8–10 years, this variability in the summer season was 6.2 ‰, in winter – 6.7 ‰ with a statistically significant difference (P > 0.95) between the spontaneous level of cytogenetic variability. The association between karyotype stability of the studied animals and seasonal environmental factors was determined using a correlation coefficient (r). A significant positive correlation was found between aneuploidy and air temperature in primiparous and cows aged 8–10 years, relative humidity, precipitation and wind strength and chromosomal gaps in primiparous and adult cows, atmospheric pressure, relative humidity and asynchronous divergence. in adult cows and cows aged 8–10 years. A positive reliable associative dependence of the appearance of micronucleus lymphocytes and dinuclear lymphocytes on the amount of precipitation and wind strength in this area in primiparous and adult cows was established.

ENDOMETRIAL STROMAL CELLS: ISOLATION, EXPANSION, MORPHOLOGICAL AND FUNCTIONAL PROPERTIES

Aim: We aimed to study biological properties of human endometrial stromal cells in vitro. Materials and Methods: The endometrium samples (n = 5) were obtained by biopsy at the first phase of the menstrual cycle from women with endometrial hypoplasia. In all cases, a voluntary written informed consent was obtained from the patients. Endometrial fragments were dissociated by enzymatic treatment. The cells were cultured in DMEM/F12 supplemented with 10% FBS, 2 mМ L-glutamine and 1 ng/ml FGF-2 in a multi-gas incubator at 5% CO2 and 5% O2. At P3 the cells were subjected to immunophenotyping, multilineage differentiation, karyotype stability and colony forming efficiency. The cell secretome was assessed by BioRad Multiplex immunoassay kit. Results: Primary population of endometrial cells was heterogeneous and contained cells with fibroblast-like and epithelial-like morphology, but at P3 the majority of cell population had fibroblast-like morphology. The cells possessed typical for MSCs phenotype CD90+CD105+CD73+CD34-CD45-HLA-DR-. The cells also expressed CD140a, CD140b, CD146, and CD166 antigents; and were negative for CD106, CD184, CD271, and CD325. Cell doubling time was 29.6 ± 1.3 h. The cells were capable of directed osteogenic, adipogenic and chondrogenic differentiation. The cells showed 35.7% colony forming efficiency and a tendency to 3D spheroid formation. The GTG-banding assay confirmed the stability of eMSC karyotype during long-term culturing (up to P8). After 48 h incubation period in serum-free medium eMSC secreted anti-inflammatory IL-1ra, as well as IL-6, IL-8 and IFNγ, angiogenic factors VEGF, GM-CSF and FGF-2, chemokines IP-10 and MCP-1. Conclusion: Thus, cultured endometrial stromal cells meet minimal ISCT criteria for MSC. Proliferative potential, karyotype stability, multilineage plasticity and secretome profile make eMSC an attractive object for the regenerative medicine use.

Karyotype stability and unbiased fractionation in the paleo-allotetraploidCucurbitagenomes

TheCucurbitagenus contains several economically important species in the Cucurbitaceae family. Interspecific hybrids betweenC. maximaandC. moschataare widely used as rootstocks for other cucurbit crops. We report high-quality genome sequences ofC. maximaandC. moschataand provide evidence supporting an allotetraploidization event inCucurbita. We are able to partition the genome into two homoeologous subgenomes based on different genetic distances to melon, cucumber and watermelon in the Benincaseae tribe. We estimate that the two diploid progenitors successively diverged from Benincaseae around 31 and 26 million years ago (Mya), and the allotetraploidization happened earlier than 3 Mya, whenC. maximaandC. moschatadiverged. The subgenomes have largely maintained the chromosome structures of their diploid progenitors. Such long-term karyotype stability after polyploidization is uncommon in plant polyploids. The two subgenomes have retained similar numbers of genes, and neither subgenome is globally dominant in gene expression. Allele-specific expression analysis in theC. maxima×C. moschatainterspecific F1hybrid and the two parents indicates the predominance oftrans-regulatory effects underlying expression divergence of the parents, and detects transgressive gene expression changes in the hybrid correlated with heterosis in important agronomic traits. Our study provides insights into plant genome evolution and valuable resources for genetic improvement of cucurbit crops.