HLA homozygous stem cell lines derived from human parthenogenetic blastocysts.

Individual HLA homozygous parthenogenetic human stem cell (hpSC-Hhom) lines have the potential for cell-based therapy in a significant number of individuals, provided the HLA haplotype is prevalent. We report the successful derivation of four stable hpSC-Hhom lines from both HLA homozygous and HLA heterozygous donors. Of these, the hpSC-Hhom-4 line carries the HLA haplotype found most commonly within the U.S. population, and is shared by different racial groups. These hpSC-Hhom lines demonstrate typical human embryonic stem cell morphology, expressing appropriate stem cell markers and possessing high levels of alkaline phosphatase and telomerase activity. Additionally, injection of these cell lines into immunodeficient animals leads to teratoma formation. G-banded karyotyping demonstrates a normal 46,XX karyotype in lines hpSC-Hhom-1 and hpSC-Hhom-4, and chromosomal anomalies in lines hpSC-Hhom-2 and hpSC-Hhom-3, both derived from the same donor. HLA genotyping of all four hpSC-Hhom lines demonstrates that they are HLA homozygous. Furthermore, in the case of HLA heterozygous donors, the hpSC-Hhom lines inherit the haplotype from only one of the donor's parents. Single-nucleotide polymorphism (SNP) data analysis suggests that hpSC-Hhom lines derived from HLA heterozygous oocyte donors are homozygous throughout the genome as assessed by SNP analysis. The protocol used for deriving these HLA homozygous stem cell lines minimizes the use of animal-derived components, which makes them more appealing for potential clinical application.

Patient-specific stem cell lines derived from human parthenogenetic blastocysts.

Parthenogenetic activation of human oocytes may be one way to produce histocompatible cells for cell-based therapy. We report the successful derivation of six pluripotent human embryonic stem cell (hESC) lines from blastocysts of parthenogenetic origin. The parthenogenetic human embryonic stem cells (phESC) demonstrate typical hESC morphology, express appropriate markers, and possess high levels of alkaline phosphatase and telomerase activity. The phESC lines have a normal 46, XX karyotype, except one cell line, and have been cultured from between 21 to 35 passages. The phESC lines form embryoid bodies in suspension culture and teratomas after injection to immunodeficient animals and give differentiated derivatives of all three embryonic germ layers. DNA profiling of all six phESC lines demonstrates that they are MHC matched with the oocyte donors. The study of imprinted genes demonstrated further evidence of the parthenogenetic origin of the phESC lines. Our research has resulted in a protocol for the production of human parthenogenetic embryos and the derivation of stem cell lines from them, which minimizes the presence of animal-derived components, making the derived phESC lines more suitable for potential clinical use.

[Plasmid DNA transfection in fibroblasts of athymic rats]. / Issledovanie transfektsii plazmidnoi DNK v fibroblasty bestimusnoi krysy.

Nontumorigenic clone FR-7 cl 13 from fibroblasts of athymic rat was obtained from stroma of human colon carcinoma xenograft propagated on nude animals. Spontaneous transformation of this cells was absent after 40 passages in vitro and treatment with pSV2neo. But cells give rise to tumors in athymic mice after transfection with pEJ. This cell clone can be recommended as cells-targets for transfection.

[Induction of stromal cell transformation in xenografts of human colonic cancer]. / Induktsiia transformatsii stromal'nykh kletok v ksenotransplantatakh raka tolstoi kishki cheloveka.

Two transformed stromal cell lines FM-7 and FR-7 were obtained from human xenografts of colon tumor (HCT-7), propagated in nude mice and nude rats, respectively. These two cell lines were confirmed to be murine and rat's by a cytogenetic study and were tumourigenic in BALB/c nude mice with the morphology of fibrosarcoma. Human DNA sequences were not detected by hybridization with pBlur8 probe in DNA of FR-7 cell line grown in vitro. These results indicate that human cancer cells from xenograft HCT-7 can induce malignancy in adjacent normal cells of nude mice and nude rats in the absence of DNA transfer.

[The evaluation of the mutagenic activity of pertussis vaccinal preparations]. / Otsenka mutagennoi aktivnosti kokliushnykh vaktsinal'nykh preparatov.

Two vaccine preparations obtained from Bordetella pertussis, whole-cell vaccine constituting one of the components of adsorbed DPT vaccine and acellular vaccine, were tested for mutagenicity. The doses of the preparations covered the range 1-100 ED50. Ames' test and the metaphase analysis of marrow cells of C57BL/6J mice were used. The acellular preparation was also tested on thymectomized mice, taking into consideration chromosomal aberrations in marrow metaphases. Whole-cell and acellular pertussis vaccines did not induce mutations in Salmonella typhimurium and chromosomal aberrations in mouse marrow cells.

[Xenograft of human RShM32 cervical cancer and comparative DNA flow cytometry]. / Geterotransplantat raka sheiki matki cheloveka RShM 32 i sravnitel'naia tsitometriia DNK.

Xenograft CC-32 was obtained from uterine cervical carcinoma of 24 years old woman in IV stage of the disease. Morphological diagnosis of clinical material and xenograft at various passages was the same-squamous cell carcinoma. Examination tumour DNA showed HPV-16 DNA integrated in five selected points. Tumor CC-32 contained a large quantity a diploid and tetraploid cells in contrast with another cervical carcinoma xenograft.

[Effect of potassium iodide on tumor growth]. / Vliianie iodida kaliia na opukholevyi rost.

In the experiments on mice with primary tumors the authors studied the effect of iodine potassium on tumoral growth. It was shown that tumors became smaller in those animals who obtained iodine potassium. Possible mechanisms of this effect are being discussed.

[Accumulation of fluorescent dyes in tumor and normal cells of man]. / Nakoplenie fliuorestsentnogo krasitelia opukholevymi i normal'nymi kletkami cheloveka.

The comparative accumulation of fluorescein Na2-salt (FINa) by the established cell lines of human tumors (cancer of the uterine body, urinary bladder, Wilms tumor, chorionepithelioma, melanoma, rhabdomyosarcoma, osteosarcoma) and human normal fibroblast cultures was obtained. The tumor cells of the different genesis is characterized by its own parameters of FINa accumulation. The most pronounced accumulation of dye has been noted for cells of cancer of the uterine body, the urinary bladder and chorionepithelioma. The normal cells accumulated FINa less considerably. Mechanism of the selective accumulation of dye by the tumor cells was discussed.

[The effect of different Yersinia pestis antigens on the cellular link in immunity]. / Izuchenie vliianiia razlichnykh antigenov Yersinia pestis na kletochnoe zveno immuniteta.

Immunization with live plague vaccine has been shown to give no protection to thymectomized mice from subcutaneous challenge with Y. pestis virulent strain. Under the action of the vaccine or individual Y. pestis antigens (fraction I) the functional and morphological activation of thymocytes and macrophages is observed, more pronounced in C57BL/6 mice and less pronounced in CBA mice. Y. pestis antigenic preparations (fractions I and II, pesticin) act as T-cell mitogens and are thus capable of inducing the in vitro proliferation of thymocytes. At the same time the in vivo action of fraction II induces a decrease in the level of lymphocytes in the peripheral blood of mice and the destruction of lymphocytes in their thymus and spleen.

[Heterografts of human cervical cancer. Characterization of papillomavirus genomes]. / Perevivaemye geterotransplantaty raka sheiki matki cheloveka. Kharakteristika genomov virusov papillomatoza.

Three new human cervical carcinoma xenografts was established from clinical tumor specimens of patients with I-II stages of disease. Growth characteristics of models are writing. Xenografts designated CC 9, 24 and 25 contain integrated DNA HPV, complemented DNA HPV-16. DNA CC 5 contain neither DNA HPV-16, nor DNA HPV-18.

[The effect of potassium iodide on the pharmacokinetics of radioactive iodine-labelled antibodies]. / Vliianie iodistogo kaliia na farmakokinetiku mechennykh radioaktivnym iodom antitel.

Radioactive iodine-labeled rabbit antibodies to beta 1-MA antigen (the marker of human colon cancer) were injected iv to BALB/c mice as well as to nude mice with xenografts of various human tumors. Preparations of iodinated antibodies contained an admixture of protein-unbound radionuclide 2-4% of total radioactivity. Part of the recipient animals were given per os 0.1% solution of KI to block the thyroid from radioactive iodine. A comparative study was made of the distribution of radioactivity in the tissues of recipients in the time course of 7 days. The thyroid of nude mice with tumors was shown to uptake much more radioactivity than that of immunologically intact mice. In addition to its protective effect of the thyroid from radioactive iodine, KI also enhanced the excretion of the preparation of labeled antibodies. Possible mechanisms of the revealed phenomena were analyzed.

[Effects of cortiphen on human tumor strains transplanted into athymic mice and rats]. / Deistvie kortifena na shtammy opukholei cheloveka, transplantiruemye bestimusnym mysham and krysam.

The antitumour activity of cortiphen synthesized at the All-Union Cancer Research Centre of the USSR Academy of Medical Sciences was studied on human tumour strains transplanted into nude mice and rats. Cortiphen was found to possess an expressed activity to kidney cancer, cancer of corpus uteri, chorionepithelioma, fibrosarcoma. Two strains of colon adenocarcinoma out of three have displayed an expressed sensitivity to cortiphen, while melanoma and Jewing sarcoma strains proved to be weakly sensitive to the preparation.