Mariusz J. Nawrocki, Piotr Celichowski, Maurycy Jankowski, Wiesława Kranc, Artur Bryja, Sylwia Borys-Wójcik, Michal Jeseta, Paweł Antosik, Dorota Bukowska, Małgorzata Bruska, Maciej Zabel, Michał Nowicki and Bartosz Kempisty
, Zabel M, Kempisty B. Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primaryculture in vitro a signaling pathways activation approach. Med J Cell Biol. 2018;6:18–26; DOI:10.2478/acb-2018-0004.
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Rossen S. Sokolov, Bistra Y. Atanassova and Elena T. Iakimova
The objective of this study was to assess the regeneration response of in vitro cultured Magnolia × soulangeana ‘Alexandrina’ and Magnolia liliiflora ‘Nigra’ to nutrient medium composition. In the primary culture (initiated from dormant axillary buds) combinations of Murashige and Skoog (MS) basal salts with 6-benzylaminopurine and α-naphthaleneacetic acid were tested. The primary explants of cv. ‘Alexandrina’ expressed higher regeneration rate than cv. ‘Nigra’. For both species, the regen eration was most strongly potentiated at addition of 0.25 mg dm−3 of the cytokinin alone. The auxin exerted undesir–able effects. Several basal salts media were applied in proliferation stage and their physiological effects were evaluated in reference to traditionally used MS. At culturing on Chée & Pool C2d Vitis Medium (VM) that is for the first time introduced to magnolia and on MS, M. liliiflora formed more but less elongated shoots than M. soulangeana. However, on VM, substantial increase (25-30%) of the number of axillary shoots and leaves, shoot length and fresh and dry weights over MS was established for both species. This suggested VM as promising composition of nutrients in multiplication stage. Microshoots obtained on MS, VM, Rugini Olive Medium and DKW Juglans Medium were successfully rooted in vitro and subsequently established ex vitro. The findings expand the information on magnolia response to culture conditions and contribute to elaboration of innovative elements of protocols for establishing tissue cultures with high regeneration capacity.
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Katarzyna Stefańska, Agata Chamier-Gliszczyńska, Maurycy Jankowski, Piotr Celichowski, Magdalena Kulus, Magdalena Rojewska, Paweł Antosik, Dorota Bukowska, Małgorzata Bruska, Michał Nowicki, Bartosz Kempisty, Michal Jeseta and Jana Zakova
The correct oviductal development and morphogenesis of its epithelium are crucial factors influencing female fertility. Oviduct is involved in maintaining an optimal environment for gametes and preimplantation embryo development; secretory oviductal epithelial cells (OECs) synthesize components of oviductal fluid. Oviductal epithelium also participates in sperm binding and its hyperactivation. For better understanding of the genetic bases that underlay porcine oviductal development, OECs were isolated from porcine oviducts and established long-term primary culture. A microarray approach was utilized to determine the differentially expressed genes during specific time periods. Cells were harvested on day 7, 15 and 30 of in vitro primary culture and their RNA was isolated. Gene expression was analyzed and statistical analysis was performed. 48 differentially expressed genes belonging to “tube morphogenesis”, “tube development”, “morphogenesis of an epithelium”, “morphogenesis of branching structure” and “morphogenesis of branching epithelium” GO BP terms were selected, of which 10 most upregulated include BMP4, ARG1, SLIT2, FGFR1, DAB2, TNC, EPAS1, HHEX, ITGB3 and LOX. The results help to shed light on the porcine oviductal development and its epithelial morphogenesis, and show that after long-term culture the OECs still proliferate and maintain their tube forming properties.
Wiesława Kranc, Maurycy Jankowski, Joanna Budna, Piotr Celichowski, Ronza Khozmi, Artur Bryja, Sylwia Borys, Marta Dyszkiewicz-Konwińska, Michal Jeseta, Magdalena Magas, Dorota Bukowska, Paweł Antosik, Klaus P. Brüssow, Małgorzata Bruska, Michał Nowicki, Maciej Zabel and Bartosz Kempisty
The ovary is part of the reproductive system, possessing very important functions in the reproduction process (ovum and embryo transfer, providing a suitable environment for sperm capacitation, etc.). There are two types of cells in the fallopian tubes: alveolar and secretive cells. These study shows the metabolic processes in pig oviductal epithelial cells associated with the activation of signaling pathways of amino acids metabolism and degradation during long-term in vitro culture. Oviductal epithelial cells from 45 colonies in the anestrous phase of the estrous cycle have been utilized in this study. RNA extract from the OEC primary cultures was pooled after 24h, 7days, 15 days and 30 days from the beginning of culture and the transcriptome investigated by Affymetrix® Porcine Gene 1.1 ST. From the whole transcript that consisted of 2009 different genes, 1537 were upregulated and 995 were downregulated after 7 days of culture, 1471 were upregulated and 1061 were downregulated after 15 days of culture and 1329 were upregulated and 1203 were downregulated after 30 days of culture. The results of these studies provide, for the first time, information on the activation of metabolic pathways of amino acids such as valine, leucine, isoleucine, cysteine, and methionine in the investigated tissue. They also indicate genes that may be OECs-specific genetic markers that are expressed or upregulated during long-term in vitro culture.
Artur Bryja, Marta Dyszkiewicz-Konwińska, Maurycy Jankowski, Piotr Celichowski, Katarzyna Stefańska, Agata Chamier-Gliszczyńska, Blanka Borowiec, Katarzyna Mehr, Dorota Bukowska, Paweł Antosik, Małgorzata Bruska, Maciej Zabel, Michał Nowicki and Bartosz Kempisty
, Kempisty B. New gene markers for metabolic processes and homeostasis in porcine buccal pouch mucosa during cells long term-cultivation - a primaryculture approach. IJMS. 2018;19(4):1027; DOI:10.3390/ijms19041027.
7. Huang DW, Sherman BT, Tan Q, Collins JR, Alvord WG, Roayaei J, Stephens R, Baseler MW, Lane HC, Lempicki RA. The DAVID Gene Functional Classification Tool: a novel biological module-centric algorithm to functionally analyze large gene lists. Genome Biol. 2007;8(9):R183; DOI:10.1186/gb-2007-8-9-r183.
8. Mering C von, Jensen LJ, Snel B, Hooper SD
Joanna Budna, Piotr Celichowski, Sandra Knap, Maurycy Jankowski, Magdalena Magas, Mariusz J. Nawrocki, Piotr Ramlau, Andrzej Nowicki, Magdalena Rojewska, Błażej Chermuła, Michal Jeseta, Paweł Antosik, Dorota Bukowska, Małgorzata Bruska, Maciej Zabel, Michał Nowicki and Bartosz Kempisty
The process of reproduction requires several factors, leading to successful fertilization of an oocyte by a single spermatozoon. One of them is the complete maturity of an oocyte, which is acquired during long stages of folliculogenesis and oogenesis. Additionally, the oviduct, composed of oviductal epithelial cells (OECs), has a prominent influence on this event through sperm modification and supporting oocyte’s movement towards uterus. OECs were isolated from porcine oviducts. Cells were kept in primary in vitro culture for 30 days. After 24h and on days 7, 15 and 30 cells were harvested, and RNA was isolated. Transcript changes were analyzed using microarrays. Fatty acids biosynthetic process and fatty acids transport ontology groups were selected for analysis and described. Results of this study indicated that majority of genes in both ontology groups were up-regulated on day 7, 15 and 30 of primary in vitro culture. We analyzed genes involved in fatty acids biosynthetic process, including: GGT1, PTGES, INSIG1, SCD, ACSL3, FADS2, FADS1, ACSS2, ALOX5AP, ACADL, SYK, ACACA, HSD17B8, FADS3, OXSM, and transport, including: ABCC2, ACSL4, FABP3, PLA2G3, PPARA, SYK, PPARD, ACACA and P2RX7. Elevated levels of fatty acids in bovine and human oviducts are known to reduce proliferation capacity of OECs and promote inflammatory responses in their microenvironment. Most of measured genes could not be connected to reproductive events. However, the alterations in cellular proliferation, differentiation and genes expression during in vitro long-term culture were significant. Thus, we can treat them as putative markers of changes in OECs physiology.
Piotr Celichowski, Karol Jopek, Marta Szyszka, Paulina Milecka, Ludwik K. Malendowicz, Marianna Tyczewska and Marcin Ruciński
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Kateřina Lněničková, Lenka Skálová, Lucie Stuchlíková, Barbora Szotáková and Petra Matoušková
10. R. Glockner, A. Lieder and A. Lupp, Determination of CYP activity in precision-cut liver slices: whether to use intact slices or slice homogenate, Anal. Bioanal. Chem. 392 (2008) 1167-1172; https://doi.org/10.1007/s00216-008-2238-y
11. M. Monshouwer, G. A. Van’t Klooster, S. M. Nijmeijer, R. F. Witkamp and A. S. van Miert, Characterization of cytochrome P450 isoenzymes in primarycultures of pig hepatocytes, Toxicol. In Vitro 12 (1998) 715-723.
12. M. N. Berry, G. J. Barritt
Ievgeniia Kocherova, Maciej Brązert, Patrycja Sujka-Kordowska, Aneta Konwerska, Magdalena Kulus, Błażej Chermuła, Piotr Celichowski, Hanna Piotrowska-Kempisty, Paweł Antosik, Dorota Bukowska, Małgorzata Bruska, Leszek Pawelczyk, Maciej Zabel, Michał Nowicki, Bartosz Kempisty and Michal Jeseta
epithelial cells (OECs) primaryculture in vitro – a signaling pathways activation approach. Med J Cell Biol. 2018;6:18–26; DOI:10.2478/acb-2018-0004. 10.2478/acb-2018-0004 Kranc W Jankowski M Budna J Celichowski P Khozmi R Bryja A Borys S Dyszkiewicz-Konwińska M Jeseta M Magas M Bukowska D Antosik P Brüssow KP Bruska M Nowicki M Zabel M Kempisty B Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primaryculture in vitro – a signaling pathways