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Clonal Cultures (clonal + culture)
Selected AbstractsPlasticity of clonal populations of dedifferentiated adult human articular chondrocytesARTHRITIS & RHEUMATISM, Issue 5 2003Andrea Barbero Objective To investigate whether adult human articular chondrocytes (AHACs), dedifferentiated by monolayer expansion, can differentiate toward diverse mesenchymal lineages and, if so, whether this ability is regulated by growth factors during monolayer expansion. Methods AHACs were expanded as multiclonal or clonal populations in medium without (control) or with factors enhancing cell dedifferentiation (transforming growth factor ,1, fibroblast growth factor 2, and platelet-derived growth factor type BB [TFP]). Cells were then cultured under conditions promoting chondrogenic, osteogenic, or adipogenic differentiation, and the acquired phenotypes were assessed histologically, biochemically, and by real-time reverse transcriptase,polymerase chain reaction. Results Multiclonal populations of both control- and TFP-expanded AHACs differentiated toward the chondrogenic, osteogenic, and adipogenic lineages. Compared with control-expanded AHACs, TFP-expanded cells displayed enhanced chondrogenic differentiation capacity (2.4-fold higher glycosaminoglycan/DNA content and 2,500-fold higher up-regulation of type II collagen) and osteogenic differentiation capacity (9.4-fold higher increase in alkaline phosphatase activity and 12.4-fold higher up-regulation of bone sialoprotein), but reduced formation of adipocytes (5.2-fold lower oil red O,positive cells/area). Clonal populations of AHACs could be efficiently expanded in TFP, but not in control medium. Most TFP-expanded clones were able to redifferentiate only into chondrocytes (7 of 20) or were unable to differentiate (6 of 20). However, some clones (2 of 20) differentiated toward all of the lineages investigated, thus displaying characteristics of mesenchymal progenitor cells. Conclusion Dedifferentiated AHACs exhibit differentiation plasticity, which is modulated by growth factors used during monolayer expansion and is highly heterogeneous across different clones. Clonal culture of AHACs in the presence of regulatory molecules could lead to the identification of AHAC subpopulations with enhanced cartilage repair capacity. [source] EFFECTS OF BARLEY STRAW EXTRACT ON GROWTH OF FIVE SPECIES OF PLANKTONIC ALGAEJOURNAL OF PHYCOLOGY, Issue 2001Article first published online: 24 SEP 200 Holz, J. C.1, Fessler, C. J.2, Severn, A. A.1 & Hoagland, K. D.1 1School of Natural Resource Sciences, University of Nebraska, 103 Plant Industry Bldg., Lincoln, NE, 68583-0814; 2Biology Department, Nebraska Wesleyan University, 5000 St. Paul Ave., Lincoln, NE, 68504; Phone: 402-472-6648; Fax: 402-472-2964 The effects of exposure to barley straw extract and the timing of exposure on the growth of four common cyanophyte species and one species of green algae were investigated in two laboratory experiments. Clonal cultures of Anabaena cylindrica, Cylindrospermum sp., Gloeocapsa sp., Eucapsis sp., and Chlorella vulgaris were obtained from culture collections. In both experiments, the algae were cultured in Guillard's WC medium at 20 °C on a 12:12 L/D photoperiod. In the first experiment, the algae were dosed with four concentrations of barley straw extract at the beginning of the experiment (day 0) and growth was monitored every second day using fluorometric detection of chlorophyll a for 14 d. In the second experiment, the algae were dosed with the same extract concentrations, but the extract was not added until the algae were in exponential growth phase (day 6). Both experiments also had control treatments (i.e. no extract) and each extract and control treatment was replicated five times. Growth of C. vulgaris was inhibited by all doses in both experiments, but inhibition was 22% greater when the extract was added on day 0. Growth Gleocapsa sp. was slightly inhibited by all doses when the extract was added on day 0, but not when it was added on day 6. No other species were inhibited, regardless of dose or timing of dose. The results of this study and other bioassay studies suggest that differential susceptibility to barley straw among algae is common and may reduce the effectiveness of barley straw as an algal control technique. [source] Pyramimonas tychotreta, sp. nov. (prasinophyceae), a new marine species from antarctica: light and electron microscopy of the motile stage and notes on growth ratesJOURNAL OF PHYCOLOGY, Issue 1 2000Niels Daugbjerg An undescribed marine prasinophyte, Pyramimonas tychotreta, sp. nov., was isolated from a water sample collected near the ice edge in the Weddell Sea (Antarctica) and is characterized by means of light and electron microscopy. This is the second described Antarctic species in the genus and it possesses a cell ultrastructure typical for members of the subgenus Vestigifera McFadden. The quadriflagellated cells measure 8,12 ,m in length and 6,7 ,m in width and are equipped with seven types of organic scales that cover the flagella and cell body. The scale floor of the box scales is ornamented by quadrants of parallel striations running perpendicular to one another. The scale floor is further characterized by a number of randomly positioned perforations. The wall of the box scales may be solid or possess up to five perforations. The base of the crown scales is square with rounded corners. It is formed of two crossed ribs, the extremities of which are interconnected by a peripheral rib. Four upright arms, attached to the peripheral rib in positions slightly offset from its junction with the cross ribs, join up with the distal extremity of a central upright strut. Each arm possesses two spines. The limuloid scales are cross-striped by 10,12 ribs. Some details of the flagellar apparatus are briefly reported. Pyramimonas tychotreta is compared with other species of the genus. Experiments were conducted to study the response of growth rate to variations in temperature and salinity in the clonal culture. The best growth rate (0.45 divisions·24 h,1) was found at 4.6° C; growth ceased at temperatures in excess of 12° C. Growth in salinities ranging from 15 to 35 psu was similar, but was arrested at 10 psu. These studies suggest that P. tychotreta as a cold stenotherm and euryhaline taxon. New observations are presented on the geographic distribution of previously described species of Pyramimonas Schmarda from the Northern Foxe Basin, Canada. [source] Exploration of the functional hierarchy of the basal layer of human epidermis at the single-cell level using parallel clonal microcultures of keratinocytesEXPERIMENTAL DERMATOLOGY, Issue 4 2010Nicolas O. Fortunel Please cite this paper as: Exploration of the functional hierarchy of the basal layer of human epidermis at the single-cell level using parallel clonal microcultures of keratinocytes. Experimental Dermatology 2010. Abstract:, The basal layer of human epidermis contains both stem cells and keratinocyte progenitors. Because of this cellular heterogeneity, the development of methods suitable for investigations at a clonal level is dramatically needed. Here, we describe a new method that allows multi-parallel clonal cultures of basal keratinocytes. Immediately after extraction from tissue samples, cells are sorted by flow cytometry based on their high integrin-,6 expression and plated individually in microculture wells. This automated cell deposition process enables large-scale characterization of primary clonogenic capacities. The resulting clonal growth profile provided a precise assessment of basal keratinocyte hierarchy, as the size distribution of 14-day-old clones ranged from abortive to highly proliferative clones containing 1.7 × 105 keratinocytes (17.4 cell doublings). Importantly, these 14-day-old primary clones could be used to generate three-dimensional reconstructed epidermis with the progeny of a single cell. In long-term cultures, a fraction of highly proliferative clones could sustain extensive expansion of >100 population doublings over 14 weeks and exhibited long-term epidermis reconstruction potency, thus fulfilling candidate stem cell functional criteria. In summary, parallel clonal microcultures provide a relevant model for single-cell studies on interfollicular keratinocytes, which could be also used in other epithelial models, including hair follicle and cornea. The data obtained using this system support the hierarchical model of basal keratinocyte organization in human interfollicular epidermis. [source] HOMOTHALLIC AUXOSPORULATION IN PSEUDO-NITZSCHIA BRASILIANA (BACILLARIOPHYTA),JOURNAL OF PHYCOLOGY, Issue 1 2009Sonia Quijano-Scheggia Most pennate diatoms are allogamous, and various types of mating systems have been described. In Pseudo-nitzschia, reproductive stages have been identified in some species, and it is generally accepted that the genus is mainly heterothallic. Here we report homothallic auxosporulation of Pseudo-nitzschia brasiliana Lundholm, Hasle et G. A. Fryxell. To our knowledge, this is the first verified description of homothallic sexual reproduction in the genus. Auxospore formation was observed in all 16 subclones derived from three initial clonal cultures of P. brasiliana. Pairing was followed by production of two gametes per gametangium, which fused to give two zygotes. Each zygote (early auxospore) was initially spherical and adhered to one girdle band of the parental frustule. The two auxospores tended to expand parallel to each other and perpendicular to the parental frustule. Elongation was synchronous, slightly asynchronous, or totally asynchronous. The entire process of sexual reproduction, from gamete formation to the appearance of the initial vegetative cells, took 2,4 d. The occurrence of sex in a homothallic species seems an advantageous life strategy for this species in that any encounter between cells of the right size class is potentially sexual. [source] Tolerance of Soil Flagellates to Increased NaCl levelsTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 4 2002FLEMMING EKELUND ABSTRACT. The ability of heterotrophic flagellates to survive and adapt to increasing salinities was investigated in this study. Whole soil samples were subjected to salinities corresponding to marine conditions and clonal cultures were used to perform growth and adaptation experiments at a wide range of different salinities (0,50 ppm). More morphotypes tolerant to elevated NaCl levels were found in road verge soil that was heavily exposed to de-icing salt than in less exposed soils, though there were fewer tolerant than intolerant morphotypes in all soils examined. Heterotrophic flagellates isolated on a freshwater medium from a non-exposed soil were unable to thrive at salinities above 15 ppt, and showed reduced growth rates even at low salt salinities (1,5 ppt). The findings suggest that heterotrophic soil flagellates are less tolerant to NaCl than their aquatic relatives, possibly due to their long evolutionary history in soil, and support the idea that identical morphospecies may differ considerably with respect to physiology [source] Use of Micromanipulation and "Feeder Layers" to Clone the Oyster Pathogen Perkinsus marinusTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 2 2000DAVID BUSHEK ABSTRACT. Genetic and biochemical characterization of microbes often requires the use of clonal cultures. A method to clone the oyster parasite Perkinsus marinus is described. Individual cells are isolated via micromanipulation and maintained above an actively proliferating "feeder layer" of P. marinus on a 0.45-,m membrane. Extracellular products released from the proliferating feeder layer can diffuse across the membrane and bathe the isolated cell, stimulating it to proliferate. The method is relatively simple and should be applicable to most protists that can be cultured in the laboratory. [source] | |||||||||||||