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PubMed1978

[Diploid and aneuploid human cell lines from autopsy material in long term cultures. Investigations of growth behaviour and karyotype (author's transl)].

Experiences with the development and characterization of cell lines obtained from human autopsy material, mainly of peripartal deceased children with multiple malformations, are reported. On 7 diploid and 4 aneuploid fibroblast-like cell lines (2 cases of trisomy G, 2 cases of trisomy E) investigations of the whole life span in vitro, on the growth behaviour and morphology as well as karyotype analyses in the early, middle and late phases of cultivation were carried out. All cell lines showed a definite limited life span, their doubling potential being on the average greater than 30 +/- 5. Compared with results from literature, these values had been found to be between those of cells of embryonic and adult origin and are possibly dependent on the age of the donor. No morphologic signs or changes of cell kinetic parameters were found which might point to a spontaneous cell alteration. The karyotype determined in the early culture phase was stable until the natural end of the cell lines. The results may serve as an evidence for the stability of all investigated parameters and for a reproducible growth behaviour in diploid and aneuploid cell lines developed from human autopsy material, a prerequisite to sue them as an in vitro model for further investigations.

Aneuploidy↗

PubMed2005

Diversity of diploid androgenic Brussels sprout plants of R0 and R1 generations.

Androgenic Brussels sprout plants were produced by the use of anther culture from the donor cultivar 'Philemon F1'. A total of 96 plants obtained from 20 androgenic R0 genotypes assigned as diploids were evaluated both in the generative and vegetative stage, in respect of their morphological characters: mean plant height; leaf size, colour and waxiness; leaf blade shape, blistering and attitude; number of sprouts; as well as their self-incompatibility and fertility. Androgenic R0 plants derived from each of the 20 embryos were highly diversified and differed from the donor in one or more morphological traits in the vegetative stage. Evaluated populations also varied in fertility and self-incompatibility. Six androgenic genotypes that set a sufficient amount of seeds of the R1 generation and 'Philemon F1' were evaluated in the field in respect of plant height, total and marketable yield per plot, shape of stem with sprouts, shape and density of sprouts, and spacing between sprouts. Only four diploid R0 and R1 populations may have some value for further breeding, as they are characterised by good vigour, high or medium ability for sprout generation, and sufficient fertility.

Brassica↗

PubMed2003

[Establishment and characteristics of the human diploid cell strain HLF-02].

OBJECTIVE: To establish the HLF-02 and provide a new material to the study of geronics, oncology, virology, pharmacology, and biogenetics. METHODS: The lung tissue was digested with 0.15% trypsin. More than 90% of the dissociated cell exhibited initial viability (trypan blue dye exclusion and the ability of adhesion). The surviving cells were analyzed by morphology, growth curves, karyotype analysis, electron microscopic observation and heterotransplantation. RESULTS: A newly established cell strain, which was designated HLF-02 had been maintained in continuous culture for over 50 generations for 10 months. Morphological observation, electron microscopic observation, karyotype analysis and heterotransplantation showed that it was corresponding to diploid cells. The doubling time of the cells was 75.5 hours. Chromosome analyses showed the human cell strain to be typical diploid appearance, and the tumorigenecity assays of nude mice were negative. CONCLUSION: The HLF-02 is a human dipolid cell strain and can serve the further studies of cell gerontics, oncology, virology, pharmacology, and biogenetics.

Cell Line↗

PubMedDate not supplied

[Obtention and characterization of a diploid cell line of human kidney].

A new diploid cell line of human embrionary kidney was obtained by serial subcultures (Mag) starting from a primary culture. It was prepared by applying the explant technique instead of the classical enzymatic digestion with trypsin. The stabilization process of the diploid line after the "secondary culture" was attained between the passes 6 and 7. 3 culture media were tested during the process. Only the MEM was completely satisfactory. The biological characteristics of the new line were as follows: fibroblastic morphology, Eagle MEM culture mean with 10% of SBF, split from 1:2 to 1:3, normal human carotype, postfreezing viability of 60%, pollutant-free and non-tumorigenic. The cryopreserved Mag subcultures 11-13 were studied from the point of view of their usefulness for the diagnosis of different viral groups.

Cell Line↗

PubMedDate not supplied

[Analysis of nucleotide sequences polymorphism of its regions of ribosomal genes of diploid Aegilops (L.) species].

rDNA sequences of ITS region including full sequences of ITS1, 5.8S and partly sequenced ITS2 of 22 accessions of 5 diploid Aegilops species, were determined. The full alignment length was 524 bp. The analysis of ITS sequences shows a number of species-specific nucleotide changes. For the first time for diploid Aegilops species an interspecific polymorphism was shown. In some positions the polymorphism within accessions was determined. It can be differences either between or within individual plants. In general both inter- and intraspecific polymorphism of ITS sequences was very low. Nucleotide polymorphism was determined only in 25 sites, 12 of which were informative.

Base Sequence↗

PubMed2005

[Mitochondrial DNA variation in diploid and triploid forms of silver crucian carp Carassius auratus gibelio].

Variation of two mtDNA fragments amplified in polymerase chain reaction was compared by the RFLP method in a population of the sympatric bisexual (diploid) and gynogenetic (triploid) forms of silver crucian carp. The mtDNA haplotypes of all individuals differed in at least 2.5% of nucleotide substitutions and fall into two phylogroups. All gynogenetic individuals have haplotypes of a single phylogroup, whereas haplotypes of diploid fish belonged to both mtDNA phylogroups. This testifies to a possibility of transformation of gynogenetic individuals into a bisexual form, whereas the reverse process is either extremely rare or impossible.

Animals↗

PubMed2005

[Cytogenetic analysis of diploid Avena L. species containing the as genome].

Cytogenetic examination showed that three diploid oat species containing the As genome are highly similar in karyotype structure and chromosome C-banding patterns. Avena strigosa is more similar to A. wiestii, while A. hirtula is to an extent separated from the two species, differing in the C-banding pattern of chromosome 6. The karyotypes of all three species harbor a small acrocentric chromosome, which is absent from diploid oat species containing other variants of the A genome. The results made it possible to assume genome specificity of the rearrangement resulting in this chromosome.

Avena↗

PubMed2005

Diploid partial mole with neonatal survival--a case report.

Partial mole is a gestational neoplastic disorder with a reported incidence of 0.005 - 0.01% of all pregnancies. The karyotype in such cases is usually triploid, baring a few exceptions. The diploid partial mole is an extremely rare entity with only few cases being documented in literature. The fetal outcome in such cases is usually poor. We describe a case of partial mole, which terminated at 28 weeks with a live diploid male fetus with good neonatal outcome. Follow-up showed no progression to malignant gestational trophoblastic disease.

Adult↗

PubMed2006

Extracting haplotypes from diploid organisms.

Each diploid organism has two alleles at every gene locus. In sexual organisms such as most plants, animals and fungi, the two alleles in an individual may be genetically very different from each other. DNA sequence data from individual alleles (called a haplotype) can provide powerful information to address a variety of biological questions and guide many practical applications. The advancement in molecular technology and computational tools in the last decade has made obtaining large-scale haplotypes feasible. This review summarizes the two basic approaches for obtaining haplotypes and discusses the associated techniques and methods. The first approach is to experimentally obtain diploid sequence information and then use computer algorithms to infer haplotypes. The second approach is to obtain haplotype sequences directly through experimentation. The advantages and disadvantages of each approach are discussed. I then discussed a specific example on how the direct approach was used to obtain haplotype information to address several fundamental biological questions of a pathogenic yeast. With increasing sophistication in both bioinformatics tools and high-throughput molecular techniques, haplotype analysis is becoming an integrated component in biomedical research.

Animals↗

PubMed1976

A perfusion culture system for virus vaccine manufacture in diploid cell cultures.

Development of a new perfusion culture system for the production of attenuated poliomyelitis virus in cultures of diploid cells is described. The growth characteristics of the diploid cells (MRC-5) were found to be normal in the perfused system. Procedures for the routine production of cell cultures at twice the cell density of stationary bottle cultures were established. The yield of virus (Lsc 2ab) per cell and per unit of surface growth area were observed to be significantly higher in the perfused system than in parallel stationary bottle culture controls--by factors of three- and five-fold, respectively. The field was confirmed to be within the experimental range when small production-scale vessels were used. Subject to satisfactory results in the testing of the virus product, this system could be highly economic in large-scale vaccine manufacture.

Cell Line↗

PubMed1991

[Comparative studies on the repetitive DNA sequences of diploid and triploid forms of Paragonimus westermani by restriction endonuclease and Southern blotting].

Restriction enzyme digestion of total genomic DNA of two chromosomes forms of Paragonimus westermani showed the presence of homologous highly repeated DNA in both diploid and triploid forms. Southern blotting analysis provided further evidence that the distribution of restriction enzyme sites (with 3 enzymes) on repetitive sequence of DNA of both forms were similar. However, with Pstl, Ddel, HaeIII and HpaII, their polymorphism revealed differences which were also found in each form tested separately with the hybridization technique. The present study, at the molecular level, supports the previously reported biological and biochemical results that they might be considered as different isolates or forms. It is suggested that PstI, DdeI, HaeIII and HpaII digestion pattern might be useful to distinguish the diploid from the triploid forms of Paragonimus westermani.

Animals↗

PubMed1990

[Study on chromosome c-banding in Paragonimus westermani: comparison of the diploid and triploid types].

A comparative study was performed in detail on the c-band pattern of diploid and triploid types of Paragonimus westermani by using BSG method. The result revealed that both types of P. westermani possessed the identical c-band pattern and c-band polymorphism. Thus the authors considered that the triploid type of P. westermani might be an autotriploid. The polymorphism of c-band pattern in diploid and the combination of homologous chromosomes with c-band polymorphism in triploid were reported for the first time.

Animals↗

PubMed1986

[Glucose metabolism and template synthesis in the mitotic cycle of human diploid fibroblasts].

The correlation between the rates of protein and nucleic acid synthesis and the activity of the key enzymes of glycolysis (hexokinase, phosphofructokinase) and pentose phosphate cycle (glucose-6-phosphate dehydrogenase) in the mitotic cycle of human diploid fibroblasts synchronized by double thymidine block was studied. It was found that the removal of the thymidine block is followed by short-term (presumably, non-specific) simultaneous stimulation of matrix syntheses, as well as by glycolytic and pentose phosphate cycle enzyme syntheses. By the beginning of the S-phase, all the processes appear to be inhibited, followed by gradual activation of glycolysis and pentose phosphate cycle reactions. The implementation of the cell cycle is concomitant with stepwise transitions of protein and hexokinase synthesis rates and ATP content to one of the following levels--basal, intermediate or maximal. Changes in the activity of glucose-6-phosphate dehydrogenase in the course of the cell cycle appear as oscillations, those in phosphofructokinase as alternative states. At stage M, the oscillatory processes are temporarily quenched, whereas the ATP content occupies an intermediate level. In contrast with diploid fibroblasts, in transformed T9 cells the enzyme activity is much higher, and the fluctuations in activity throughout the cell cycle are less noticeable. Presumably, in transformed cells the enzyme activity is at the maximum level and is not prone to effector regulation.

Cells, Cultured↗

PubMed1988

[RNA and protein content in cells of diploid and tetraploid toads of the Bufo viridis group].

Nucleic acids and total protein contents in hepatocytes and erythrocytes of diploid (Bufo viridis, 2n = 22) and tetraploid (B. danatensis, 2n = 44) green toads were measured cytophotometrically. The RNA (table 1) and protein (table 2) contents in the tetraploid cells are twice as much as that in diploid ones. According to these results tetraploidy in green toads may have a recent origin. The relation between structural and functional polyploidization in tetraploid amphibian species is discussed.

Animals↗

PubMed1989

[Dependence of lethality and incidence of chromosome aberrations induced by treatment of synchronized human diploid fibroblasts with sodium fluoride on different periods of the cell cycle].

The cytotoxic and clastogenic effects of sodium fluoride during various phases of cell cycle of human cultured diploid fibroblasts were examined. The cells in confluence were synchronized at G1/G0 phase by a period of growth in medium containing 1% serum (low serum medium). To obtain the cells in S phase and G2 phase, exponentially growing cells were cultured in low serum medium with a subsequent blockage of the cells at the G1/S boundary by hydroxyurea. Synchronized cells were treated with sodium fluoride for 3 hr during the G1 phase or G2 phase, and for each of three 3-hr periods during the S phase which lasted 9 hr. The cytotoxicity, as determined by a decrease in colony-forming ability, was dependent upon the phase of cell cycle during which treatment was administered. The highest lethality was observed for treatment in early to middle S phase, whereas scare lethality was observed in G1 phase. Inducibility of chromosome aberrations of the cells following treatment with sodium fluoride was also dependent upon the phase of cell cycle. Significant increase in the incidence of chromosome aberrations was observed only in cultures treated during early and/or middle S phases of the cell cycle. These results indicate that cytotoxicity and clastogenicity of sodium fluoride to cultured human diploid fibroblasts are cell phase dependent, and that the cells in early and middle S phases are more sensitive to these effects.

Cell Cycle↗

PubMed1989

[Ultrastructural studies on the spermatogenesis in diploid and triploid types of Paragonimus westermani].

Diploid and triploid types of Paragonimus westermani were compared in the aspect of ultrastructure of spermatogenesis. The results showed that the process of spermatogenesis in the diploid type was normal. The cells at every stage are well developed. A great number of morphologically and functionally normal spermatozoa could be formed. On the other hand, the process of spermatogenesis in the triploid type was abnormal, the cells at every stage developed abnormally. They were apparently degenerated or vacuolated, lacking organelles. The cytophore was broken into small particles. No normal spermatozoa were found (Figs. 1-5).

Animals↗

PubMed1989

[Microheterogeneity of the primary structure of the short repeated 5S DNA unit in diploid wheat Triticum monococcum L].

Nucleotide sequences of two 5S rRNA genes located in repeated 327 bp long units were determined in diploid wheat Triticum monococcum. They were compared with sequences of 5S rRNA genes of Tr. monococcum and Tr. aestivum which were earlier determined. The differences were revealed in two localizations of the nucleotide sequence in 5S DNA coding regions of Tr. monococcum and - in nine localizations in nontranscribed spacer. It was established that the nucleotide sequence of 5S rRNA gene cloned in pTm5S9 plasmid and 5S DNA coding region in Tr. aestivum have significant homology. Diploid wheat Tr. monococcum was supposed to have 5S rRNA genes with different functional activity within one multigene family.

Base Sequence↗

PubMedDate not supplied

[Primary structure of the 5.8S gene of rRNA and internal transcribed spacers of rDNA in diploid wheat Triticum urartu thum. ex. gandil].

Nucleotide sequences of 5.8S rRNA gene and rDNA internal transcribed spacers ITS-1 and ITS-2 were determined in diploid wheat Triticum urartu. It was shown that 5.8S rRNA gene of this wheat species consists of 163 base pairs and GC-content is 59.5%. When comparing 5.8S rRNA sequences in diploid wheat, rice and lupine and also 5.8S rRNA in hexaploid wheat and horse beans a high evolutional conservatism of its structure was revealed. The size of ITS-1 and ITS-2 in Tr. urartu is 219 and 225 base pairs long correspondingly. While comparing structures of similar rDNA regions of Tr. urartu, rice and maize a high level of homology was found only between nucleotides adjoining genes of high molecular rRNAs. In ITS-1 of Tr. urartu an insertion of 5'-GACGACGACATTGTCCGTC-3' was found, which is absent in maize and rice.

Base Sequence↗