scholarly journals Tissue culture of Papaver bracteatum. I. Cultural conditions on growth of the cell culture of Papaver bracteatum.

1976 ◽  
Vol 40 (5) ◽  
pp. 899-906 ◽  
Author(s):  
Shoji KAMIMURA ◽  
Mie AKUTSU
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Papaver Bracteatum ◽  
Cultural Conditions

Cytogenetic abnormalities of cotton somaclones from callus cultures

Genome ◽  
1989 ◽  
Vol 32 (5) ◽  
pp. 762-770 ◽  
Author(s):  
David M. Stelly ◽  
D. W. Altman ◽  
R. J. Kohel ◽  
T. S. Rangan ◽  
E. Commiskey
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Somaclonal Variation ◽  
High Frequency ◽  
Chromosomal Abnormalities ◽  
Callus Cultures ◽  
Meiotic Abnormalities ◽  
High Frequencies ◽  
Field Environment ◽  
Metaphase I

Somaclonal variation occurs among regenerants from tissue culture of many plant species. Our objective was to determine whether cytogenetic variation contributes to somaclonal variation in cotton (Gossyptum hirsutum L.,2n = 4x = 52). Of 117 somaclones of cotton regenerated from 18-month-old callus cultures of 'SJ-2' and 'SJ-5' cultivars, 35 were analyzed for meiotic abnormalities. The population of somaclones was extremely varied in phenotype, most plants being strikingly aberrant in phenotype. Fertility was generally poor: 84% failed to set bolls and only 5% set 10 or more bolls in a field environment. Only one of the somaclones (3%) formed 26 bivalents at metaphase I. Fourteen were nonsynaptic to partially synaptic at metaphase I. Synaptic abnormalities impaired fertility and precluded thorough metaphase analysis. Chromosome numbers obtained for 32 plants ranged from 49 to 53, and only 1 plant was hyperaneuploid. No plant was polyploid. Chromosomal abnormalities in plants with normal metaphase pairing included univalents, unequal bivalents, rod bivalents, trivalents, open quadrivalents, and centric fragments. Seventeen hypoaneuploid plants formed a V-shaped trivalent at metaphase I, constituting a high frequency of tertiary monosomy. The high frequencies of aneuploidy and tertiary monosomy indicate that cytogenetic anomalies are a major source of somaclonal variation in cotton. It is hypothesized that (i) primary cytogenetic events during cotton cell culture give rise to breakage – fusion – bridge (BFB) cycles, (ii) BFB cycles accrue during culture, (iii) BFB cycles cause loss of chromatin, and (iv) BFB cycles are resolved by the formation of stable tertiary chromosomes with mono-centric activity. The hypothesis accounts mechanistically for the coincidence of chromatin deficiencies and chromatin exchange involved implicitly in tertiary monosomy, as well as for the relatively high frequency of tertiary monosomy among somaclones.Key words: aneuploid, monosomic, synaptic, sterility, Gossypium.

scholarly journals Mechanism of Cell Culture Adaptation of an Enteric Calicivirus, the Porcine Sapovirus Cowden Strain

2015 ◽  
Vol 90 (3) ◽  
pp. 1345-1358 ◽  
Author(s):  
Zhongyan Lu ◽  
Masaru Yokoyama ◽  
Ning Chen ◽  
Tomoichiro Oka ◽  
Kwonil Jung ◽  
...  
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Amino Acid ◽  
Virus Replication ◽  
Amino Acid Substitutions ◽  
Virus Binding ◽  
Culture Adaptation ◽  
Porcine Sapovirus ◽  
Cowden Strain

ABSTRACTThe porcine sapovirus (SaV) (PoSaV) Cowden strain is one of only a few culturable enteric caliciviruses. Compared to the wild-type (WT) PoSaV Cowden strain, tissue culture-adapted (TC) PoSaV has two conserved amino acid substitutions in the RNA-dependent RNA polymerase (RdRp) and six in the capsid protein (VP1). By using the reverse-genetics system, we identified that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328), but not the substitutions in the RdRp region, were critical for the cell culture adaptation of the PoSaV Cowden strain. The other two substitutions in VP1 (residues 291 and 295) reduced virus replicationin vitro. Three-dimensional (3D) structural analysis of VP1 showed that residue 178 was located near the dimer-dimer interface, which may affect VP1 assembly and oligomerization; residues 289, 291, 324, and 328 were located at protruding subdomain 2 (P2) of VP1, which may influence virus binding to cellular receptors; and residue 295 was located at the interface of two monomeric VP1 proteins, which may influence VP1 dimerization. Although reversion of the mutation at residue 291 or 295 from that of the TC strain to that of the WT reduced virus replicationin vitro, it enhanced virus replicationin vivo, and the revertants induced higher-level serum and mucosal antibody responses than those induced by the TC PoSaV Cowden strain. Our findings reveal the molecular basis for PoSaV adaptation to cell culture. These findings may provide new, critical information for the cell culture adaptation of other PoSaV strains and human SaVs or noroviruses.IMPORTANCEThe tissue culture-adapted porcine sapovirus Cowden strain is one of only a few culturable enteric caliciviruses. We discovered that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328) were critical for its adaptation to LLC-PK cells. Two substitutions in VP1 (residues 291 and 295) reduced virus replicationin vitrobut enhanced virus replication and induced higher-level serum and mucosal antibody responses in gnotobiotic pigs than those induced by the tissue culture-adapted strain. Structural modeling analysis of VP1 suggested that residue 178 may affect VP1 assembly and oligomerization; residues 289, 291, 324, and 328 may influence virus binding to cellular receptors; and residue 295 may influence VP1 dimerization. Our findings will provide new information for the cell culture adaptation of other sapoviruses and possibly noroviruses.

scholarly journals HURLER'S SYNDROME

1966 ◽  
Vol 123 (1) ◽  
pp. 1-16 ◽  
Author(s):  
B. Shannon Danes ◽  
Alexander G. Bearn
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Alcian Blue ◽  
X Chromosome ◽  
Toluidine Blue ◽  
Normal Subjects ◽  
Clinically Normal ◽  
Hurler’S Syndrome ◽  
Metachromatic Granules ◽  
Primary Action

Seven families affected with Hurler's syndrome have been studied using the methods of cell culture. Skin fibroblasts obtained from the skin of 7 patients with Hurler's syndrome contained metachromatic granules when stained for mucopolysaccharides with toluidine blue O and alcian blue, whereas fibroblasts from normal subjects contained no metachromatic granules. In four families skin cultures of the clinically normal parents showed fibroblasts which contained demonstrable metachromatic granules and "gargoyle" cells and were considered to be heterozygous for the abnormal gene. Fibroblast cultures from certain other members of these families showed metachromasia. These findings were also considered to indicate heterozygosity for the abnormal gene. Three families of the X-linked type of the disease were studied. Fibroblasts cultured from the father contained no metachromatic granules whereas those of the hemizygous mother contained both metachromatic granules and "gargoyle" cells. In one family the abnormal gene could be traced through unaffected individuals for three generations. The prolonged preservation of the biochemical trait in tissue culture will permit studies to be performed designed to clarify the primary action of the abnormal genes which result in Hurler's syndrome, as well as to increase the usefulness of this trait in mapping the human X chromosome.

Patterns of metastatic (met) disease sites in breast cancer (BrCa): Implications for availability of fresh tumor tissue (FTT) for personalized BrCa treatment (Rx) planning (TP) in met disease

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e12004-e12004
Author(s):  
C. A. Presant ◽  
L. Bosserman ◽  
F. Howard ◽  
B. Emilio
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Tissue Culture ◽  
Lymph Node ◽  
Clinical Characteristics ◽  
Tumor Tissue ◽  
Cancer Center ◽  
Archival Tissue ◽  
Non Invasive ◽  
Video Assisted

e12004 Background: Personalized Rx for patients (PTs) can improve response. Although archival tissue is available in all PTs, FTT testing of molecular, immunohistochemical, and/or chemosensitivity characteristics may be more optimal for TP than tissues obtained before previous Rx and/or new mets, and archival tissue is inadequate for cell culture information. This study was performed to determine the availability of FTT in a multi-site cancer center. Methods: Pts seen over 22 months were evaluated. PT clinical characteristics were evaluated to determine the prevalence of mets by site, and feasibility of biopsy (bx) for FTT by site. A bx algorithm for FTT was developed and quantified. Results: 26,794 successive PTs were evaluated. BrCa was present in 2043 (7.6%). Median age was 61 (range 27 to 98). Mets were present in only 174 (8.5% of BrCa) who had 256 sites involved (1.47 site per PT). 68% of PTs had multiple sites involved. Of PTs with multiple sites, 96 (81%) had 2, 16 (14%) had 3, and 6 (5%) had 4 sites involved. Single site was the pattern in only 9% of liver, 7% of lung, 32% of bone, 0% of adrenal 6% of skin, but 71% of brain mets. The most frequent sites of mets were bone (50%), lung (31%), liver (26%), brain (10%), skin or soft tissue (11%), lymph node (6%), and effusion/ascites (3.4%). In order to theoretically obtain FTT for development of a personalized TP based on an algorithm preferring in order local bx > para/thoracentesis, > non-osseous needle bx > video-assisted tissue bx > osseous bx, 18% of PTs would have had a local bx, 3% para/thoracentesis, 23% liver bx, 19% lung bx, 17% bone bx, and 12% would not have had a bx because of brain-only mets. Conclusions: Most BrCa PTs in oncology practices have only locoregional disease. Most PTs with mets have multiple sites. FTT acquisition would be feasible for 88% of PTs with mets for personalized TP, and in most the bx would be simple and non-invasive. However, tissue culture tests would necessitate a more invasive video-assisted bx in 42% of PTs. Physicians should consider additional bx to personalize TP when appropriate for improving outcomes. [Table: see text]

Laminin-511 and recombinant vitronectin supplementation enables human pluripotent stem cell culture and differentiation on conventional tissue culture polystyrene surfaces in xeno-free conditions

2021 ◽  
Author(s):  
Ya-Chu Liu ◽  
Lee-Kiat Ban ◽  
Henry Hsin-Chung Lee ◽  
Hsin-Ting Lee ◽  
Yu-Tang Chang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Extracellular Matrix ◽  
Tissue Culture ◽  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Culture Conditions ◽  
Human Pluripotent Stem Cell ◽  
Tissue Culture Polystyrene ◽  
Ecm Proteins

Human pluripotent stem cells (hPSCs) are typically cultivated on extracellular matrix (ECM) protein-coated dishes in xeno-free culture conditions. We supplemented mixed ECM proteins (laminin-511 and recombinant vitronectin, rVT) in culture...

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