New Molecular Markers Involved in Regulation of Ovarian Granulosa Cell Morphogenesis, Development and Differentiation during Short-Term Primary In Vitro Culture-Transcriptomic and Histochemical Study Based on Ovaries and Individual Separated Follicles

被引:15
作者
Kulus, Magdalena [1 ]
Sujka-Kordowska, Patrycja [2 ]
Konwerska, Aneta [2 ]
Celichowski, Piotr [2 ]
Kranc, Wieslawa [3 ]
Kulus, Jakub [1 ]
Piotrowska-Kempisty, Hanna [4 ]
Antosik, Pawel [1 ]
Bukowska, Dorota [1 ]
Izycki, Dariusz [5 ]
Bruska, Malgorzata [3 ]
Zabel, Maciej [6 ,7 ]
Nowicki, Michal [2 ]
Kempisty, Bartosz [2 ,3 ,8 ,9 ]
机构
[1] Nicolaus Copernicus Univ Torun, Vet Ctr, PL-87100 Torun, Poland
[2] Poznan Univ Med Sci, Dept Histol & Embryol, PL-61701 Poznan, Poland
[3] Poznan Univ Med Sci, Dept Anat, PL-61701 Poznan, Poland
[4] Poznan Univ Med Sci, Dept Toxicol, PL-61701 Poznan, Poland
[5] Poznan Univ Med Sci, Dept Canc Immunol, Chair Biotechnol, PL-61701 Poznan, Poland
[6] Wroclaw Med Univ, Dept Histol & Embryol, PL-50367 Wroclaw, Poland
[7] Univ Zielona Gora, Div Anat & Histol, PL-65417 Zielona Gora, Poland
[8] Univ Hosp, Dept Obstet & Gynecol, Brno 60177, Czech Republic
[9] Masaryk Univ, Brno 60177, Czech Republic
关键词
pig; granulosa; cells morphogenesis; PORCINE; IDENTIFICATION; GENE; PROLIFERATION; GROWTH;
D O I
10.3390/ijms20163966
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nowadays, science has a lot of knowledge about the physiology of ovarian processes, especially folliculogenesis, hormone production and ovulation. However, the molecular basis for these processes remains largely undiscovered. The cell layer surrounding the growing oocyte-granulosa cells-are characterized by high physiological capabilities (e.g., proliferation, differentiation) and potential for growth in primary cultures, which predisposes them for analysis in the context of possible application of their cultures in advanced methods of assisted reproduction. In this study, we have used standard molecular approaches to analyze markers of these processes in primarily in vitro cultured porcine granulosa, subjected to conditions usually applied to cultures of similar cells. The material for our research came from commercially slaughtered pigs. The cells were obtained by enzymatic digestion of tissues and in vitro culture in appropriate conditions. The obtained genetic material (RNA) was collected at specific time intervals (0 h-before culture; reference, 48, 98, 144 h) and then analyzed using expression microarrays. Genes that showed a fold change greater than |2| and an adjusted p value lower than 0.05 were described as differentially expressed. Three groups of genes: Cell morphogenesis, cell differentiation and cell development were analyzed. From 265 differently expressed genes that belong to chosen ontology groups we have selected DAPL1, CXCL10, NEBL, IHH, TGFBR3, SCUBE1, DAB1, ITM2A, MCOLN3, IGF1 which are most downregulated and PDPN, CAV1, TMOD1, TAGLN, IGFBP5, ITGB3, LAMB1, FN1, ITGA2, POSTN genes whose expression is upregulated through the time of culture, on which we focused in downstream analysis. The results were also validated using RT-qPCR. The aim of our work was to conduct primary in vitro culture of granulosa cells, as well as to analyze the expression of gene groups in relation to the proliferation of follicular granulosa cells in the model of primary culture in real time. This knowledge should provide us with a molecular insight into the processes occurring during the in vitro cultures of porcine granulosa cells, serving as a basic molecular entry on the extent of the loss of their physiological properties, as well as gain of new, culture-specific traits.
引用
收藏
页数:18
相关论文
共 45 条
  • [1] Antosik, 2015, AUSTIN J INVIT FERTI, V2, P1023
  • [2] Borys-Wojcik S., 2018, MED J CELL BIOL, V6, P155, DOI [10.2478/acb-2018-0025, DOI 10.2478/acb-2018-0025, DOI 10.2478/ACB-2018-0025]
  • [3] Budna J., 2018, MED J CELL BIOL, V6, P48, DOI [10.2478/acb-2018-0009, DOI 10.2478/ACB-2018-0009]
  • [4] Analysis of proteomic changes induced upon cellular differentiation of the human intestinal cell line Caco-2
    Buhrke, Thorsten
    Lengler, Imme
    Lampen, Alfonso
    [J]. DEVELOPMENT GROWTH & DIFFERENTIATION, 2011, 53 (03) : 411 - 426
  • [5] Chamier-Gliszczyska A., 2018, Med. J. Cell Biol, V6, P163, DOI [10.2478/acb-2018-0026, DOI 10.2478/ACB-2018-0026, DOI 10.2478/acb-2018-0026]
  • [6] New Gene Markers of Angiogenesis and Blood Vessels Development in Porcine Ovarian Granulosa Cells during Short-Term Primary Culture In Vitro
    Chermula, Blazej
    Brazert, Maciej
    Izycki, Dariusz
    Ciesiolka, Sylwia
    Kranc, Wieslawa
    Celichowski, Piotr
    Ozegowska, Katarzyna
    Nawrocki, Mariusz J.
    Jankowski, Maurycy
    Jeseta, Michal
    Antosik, Pawel
    Bukowska, Dorota
    Skowronski, Mariusz T.
    Brussow, Klaus P.
    Bruska, Malgorzata
    Pawelczyk, Leszek
    Zabel, Maciej
    Nowicki, Michal
    Kempisty, Bartosz
    [J]. BIOMED RESEARCH INTERNATIONAL, 2019, 2019
  • [7] Influence of Estradiol-17beta on Progesterone and Estrogen Receptor mRNA Expression in Porcine Follicular Granulosa Cells during Short-Term, In Vitro Real-Time Cell Proliferation
    Ciesiolka, Sylwia
    Budna, Joanna
    Jopek, Karol
    Bryja, Artur
    Kranc, Wieslawa
    Chachula, Adrian
    Borys, Sylwia
    Konwinska, Marta Dyszkiewicz
    Ziolkowska, Agnieszka
    Antosik, Pawel
    Bukowska, Dorota
    Brussow, Klaus P.
    Bruska, Malgorzata
    Nowicki, Michal
    Zabel, Maciej
    Kempisty, Bartosz
    [J]. BIOMED RESEARCH INTERNATIONAL, 2016, 2016
  • [8] Transcriptional Profiling of Cultured, Embryonic Epicardial Cells Identifies Novel Genes and Signaling Pathways Regulated by TGFβR3 In Vitro
    DeLaughter, Daniel M.
    Clark, Cynthia R.
    Christodoulou, Danos C.
    Seidman, Christine E.
    Baldwin, H. Scott
    Seidman, J. G.
    Barnett, Joey V.
    [J]. PLOS ONE, 2016, 11 (08):
  • [9] Dyszkiewicz-Konwinska M, 2017, J BIOL REG HOMEOS AG, V31, P855
  • [10] Angiogenesis in the corpus luteum
    Hamish M Fraser
    Christine Wulff
    [J]. Reproductive Biology and Endocrinology, 1 (1)