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Retroviral infection of chicken testicular cells in a process of construction of transgenic poultry
Kalina, Jiří ; Škvor, Jiří (advisor) ; Petr, Jaroslav (referee) ; Jílek, František (referee)
Biotechnological research in chicken transgenesis is still lagging beyond the mammals mainly due to the specificities of avian reproductive system. This thesis is trying to offer functionally complex approach to the chciken transgenesis through one of the techniques. Common transfection techniques were applied on chicken blastodermal and testicular cells to affirm this approach. Our original techniqe of sterilization of chicken testes was improved and applied. Stained testicular cells of donor male were transplanted into sterilized acceptors testes and subseqent tubuli recolonization and sperm production were described. The retroviral- based vector was developed and transplanted testicular cells were successfuly infected. Carried marker transgene (Green fluorescence protein, GFP) was detected in DNA of produced sperms and no significant CpG methylation was detected when sreening infected cells. Through the flow cytometry, testicular cells used for transplantation were analyzed. All major spermatogonia-derived populations were described including the side population (SP), possible group of spermatogonial stem cells.
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Immunologic and epigenic modulation of gene expression of human leukemia cell lines
Elknerová, Klára ; Stöckbauer, Petr (advisor) ; Špíšek, Radek (referee) ; Škvor, Jiří (referee)
Therapy by monoclonal antibody to CD34 molecule Growth-inhibitory and proapoptotic effect of the monoclonal antibody to CD34 molecule, clone 4H11, were tested on CD34+ leukemic cell lines (MOLM-9, JURL-MK1) and CD34- cell line (PS-1). We have found that the monoclonal antibody to CD34 inhibited the proliferation and induced apoptosis of all CD34+ cell lines tested, however it has similar effect on CD34- leukemic cell line (PS-1) in concentration higher than 32 µg/ml. We did not observe induction of differentiation by anti-CD34 antibody, but a growth arrest of cells in the G0/G1 phase of the cell cycle was detected in CD34+ cell lines. Combinations of anti-CD34 antibody with both type I (α, β) or type II ( ) interferons did not enhance the effects on the cell growth or inhibition of cellular proliferation of the antibody alone. Combinations of anti-CD34 antibody with hematopoietic cytokines or INF-γ did not induced diferentiation. Our data suggest that the monoclonal antibody to CD34 molecule prepared from clone 4H11, after sufficient experimental and preclinical testing on laboratory animals, may provide a new basis for possible targeted antibody therapy of acute or chronic myeloid leukemia. Therapy by histone deacetylase inhibitors Histone deacetylase inhibitors (HDACi) are emerging new class of...
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