Katarzyna Stefańska, Ievgenia Kocherova, Sandra Knap, Magdalena Kulus, Piotr Celichowski and Michal Jeseta
fertilization, our aim was to gain a better insight into genetic changes that underlie these cells’ function. We established a long-term primaryculture of porcine OECs and performed microarray analysis, checking gene expression in specific time periods. Our results revealed a set of genes belonging to “maintenance of location”, “maintenance of protein location in cell” and “maintenance of protein location” that were differentially expressed, indicating that they may be possible marker genes of biochemical and morphological changes in OECs.
Material and Methods
Wiesława Młodawska, Patrycja Mrowiec, Beata Grabowska, Joanna Waliszewska, Joanna Kochan, Agnieszka Nowak, Anna Migdał, Wojciech Niżański, Sylwia Prochowska, Agnieszka Partyka, Marcin Pałys, Teresa Grega and Józef Skotnicki
Kanazawa S., Fujiwara T., Matsuzaki S., Shingaki K., Taniguchi M., Miyata S., Tohyama M., Sakai Y., Yano K., Hosokawa K., Kubo T. (2010). bFGF regulates PI3-kinase-Rac1-JNK pathway and promotes fibroblast migration in wound healing. PLoS One, 5(8): e12228.
Kim G.A., Oh H.J., Kim M.J., Jo Y.K., Choi J., Kim J.W., Lee T.H., Lee B.C. (2015). Effect of primaryculture medium type for culture of canine fibroblasts on production of cloned dogs. Theriogenology, 84: 524–530.
Lee H.S., Yu X.F., Bang J.I., Cho S.J., Deb G.K., Kim B.W., Kong I
Jeanne Adiwinata Pawitan, Isabella Kurnia Liem, Des Suryani, Arleni Bustami and Reza Yuridian Purwoko
Background: Lipoaspirate contains noxious substances derived from liposuction. Therefore, extensive washing is recommended before the lipoaspirate is processed further for culture or fat grafting. Washing a small amount of lipoaspirate may not pose a problem, but washing a large volume of lipoaspirate may be cumbersome, time consuming, and requires a lot of phosphate buffered saline (PBS).
Objective: To introduce a simple method for lipoaspirate washing using fine-mesh stainless-steel tea or coffee filter, a small tea spoon, and a porcelain bowl.
Methods: The filter was used to collect the adipose tissue fragments. Further washing of the fragments was achieved by soaking the adipose tissue containing filter in a PBS containing porcelain bowl and stirring using a small tea spoon to transfer the contaminating materials to the PBS. Enzymatic processing to dissociate the cells from the tissue and primary cultures was conducted as usual in MesenCult.
Results: Using the equipment mentioned above, the adipose tissue fragments were readily separated from the blood, free lipids, anesthetics, and other noxious material in the liquid portion. This simple method saves time and PBS compared with previously described methods. Further enzymatic processing produced sufficient cells to be cultured, and culture results showed plastic adherent cells on day 2 that became confluent on day 6.
Conclusion: Lipoaspirate washing using a fine mesh stainless steel filter is time saving and produced cells that grow well in MesenCult.
Oxygen (O2) is an essential element for aerobic respiration. Atmospheric concentration of O2 is approximately 21%. Mammalian cells, however, are generally adapted to O2 levels much lower than atmospheric conditions. The pericellular levels of O2 must also be maintained within a fairly narrow range to meet the demands of cells. This applies equally to cells in vivo and cells in primary cultures. There has been growing interest in the performance of cell culture experiments under various O2 levels to study molecular and cellular responses. To this end, a range of technologies (e.g. gas-permeable technology) and instruments (e.g. gas-tight boxes and gas-controlled incubators) have been developed. It should be noted, however, that some of these have limitations and they are still undergoing refinement. Nevertheless, better results should be possible when technical concerns are taken into account. This paper aims to review various aspects of O2 level adjustment in primary cell cultures, regulation of pericellular O2 gradients and possible effects of the cell culture medium.
Yu Chen, Li-ming Fu, Xiu-ying Zhao, Jun Zhao and Zhong-ping Duan
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3 Shi Q, Gaylor JD, Cousins R, Plevris J, Hayes PC, Grant MH. The effects of plasma from patients with acute liver failure on the growth and metabolism of HepG2
The Effect of Laurel Leaf Extract Against Toxicity Induced by 2,3,7,8-Tetrachlorodibenzo-P-Dioxin in Cultured Rat Hepatocytes
2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a very toxic environmental pollutant that raises great public concern about its impact on human health. Recent studies indicate that laurel leaf extract exhibits antioxidant properties that can counter the toxic effects of certain compounds in the liver. The aim of this study was to assess how effective LE is against the toxicity of TCDD in a primary culture of rat hepatocytes. The extract (50 mg L-1, 100 mg L-1, and 200 mg L-1) was added to cultures alone or with TCDD (1.61 mg L-1 and 3.22 mg L-1) for 48 hours. Cell viability was measured using the [3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide] (MTT) assay and the lactate dehydrogenase (LDH) cytotoxicity assay, while oxidative damage was assessed by measuring total antioxidant capacity (TAC) and total oxidative stress (TOS). DNA damage was also analysed using the micronucleus (MN) assay of the cultured hepatocytes. TCDD alone lowered, and laurel extract had no effect on cell viability. TCDD also increased TOS and significantly decreased TAC. It significantly increased the frequency of micronucleated hepatocytes in a dose-dependent manner. In cultures exposed to LE alone, TOS did not change and TAC significantly increased in a dose-dependent manner. Added to TCDD, laurel countered its toxic effects and showed protective effects against TCDD-mediated DNA damage. This points to the therapeutic potential of laurel against TCDD toxicity in the liver.
Artur Bryja, Marta Dyszkiewicz-Konwińska, Maurycy Jankowski, Piotr Celichowski, Katarzyna Stefańska, Agata Chamier-Gliszczyńska, Małgorzata Popis, Katarzyna Mehr, Dorota Bukowska, Paweł Antosik, Małgorzata Bruska, Maciej Zabel, Michał Nowicki and Bartosz Kempisty
real-time long-term cell proliferation in vitro: A primaryculture approach. J Biol Regul Homeost Agents. 2017;31(3):567–77.
3. Dyszkiewicz-Konwińska M, Bryja A, Jopek K, Budna J, Khozmi R, Jeseta M, Bukowska D, Antosik P, Bruska M, Nowicki M, Zabel M, Kempisty B. Expression of genes responsible for cell morphogenesis involved in differentiation in porcine buccal pouch mucosal cells during long-term primaryculture and real-time proliferation in vitro. J Biol Regul Homeost Agents. 2017;31(4):855–64.
4. Bryja A, Dyszkiewicz-Konwińska M, Budna J, Ciesiółka
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8. Moody, D. E., Fang Lin, S. N., Weyant, D. M., Strom, S. C., Omiecinski, C. J., 2009: Effect of Rifampin and Nelfinavir on the metabolism of methadone and buprenorphine in primarycultures of human hepatocytes. Drug Metab. Dispos., 37, 2323-2329.
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I. Hodorova, J. Mihalik, J. Vecanova, M. Dankova and S. Rybarova
primarycultures of human proximal tubular cells. Toxicology 2008; 244:56-65.
de Zwart L, Scholten M, Monbaliu JG, Annaert PP, Van Houdt JM, Van den Wyngaert I, De Schaepdrijver LM, Bailey GP, Coogan TP, Coussement WC, Mannens GS. The ontogeny of drug metabolizing enzymes and transporters in the rat. Reprod Toxicol. 2008, 26(3-4):220-230.
Rosati A, Maniori S, Decorti G, Candussio L, Giraldi T, Bartoli F. Physiological regulation of P-glycoprotein, MRP1, MRP2 and cytochrome P450 3A2 during rat ontogeny. Dev Growth
David Calderón Guzmán, Norma Osnaya Brizuela, Maribel Ortíz Herrera, Ernestina Hernández García, Gerardo Barragán Mejía, Hugo Juárez Olguín, Armando Valenzuela Peraza, Jonas Attilus and Norma Labra Ruíz
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3. M. Hartbauer, B. Hutter-Paie and G. Skofitsch, Antiapoptotic effects of the peptidergic drug cerebrolysin on primarycultures of embryonic chick cortical neurons, J