Growth and metabolism of Ricinus communis endosperm in tissue culture

1970 ◽  
Vol 48 (12) ◽  
pp. 2323-2331 ◽  
Author(s):  
D. J. Brown ◽  
D. T. Canvin ◽  
B. F. Zilkey
Keyword(s):  
Tissue Culture ◽  
Ricinus Communis ◽  
Glyoxylate Cycle ◽  
Culture Conditions ◽  
Endosperm Tissue ◽  
Ricinus Communis L ◽  
Culture Growth ◽  
Germinating Seeds

Endosperm tissue from both developing and germinating castor oil seeds (Ricinus communis L.) was grown as a callus in tissue culture. Callus growths were established from endosperm explants (without embryo) at all stages of seed development except from the quiescent unimbibed seed. Best tissue culture growth was observed with endosperm tissue obtained from seeds that had been germinated for 2 days.The lipid reserves diminished in all endosperm tissue which had been established on culture from germinating seeds but the rate of breakdown was much slower in cultures from 0-day germinated seeds. Glyoxylate cycle activity, as tested by acetate-1- and -2-14C incorporation, was not evident in long-term cultures.The process of lipid accumulation found in normal developing endosperm (24 to 36 days after fertilization) was not retained in culture but was replaced by a process resulting in the rapid loss of the already accumulated oil reserves.As far as could be determined the in vivo biochemical capabilities of the castor bean endosperm were not retained for a lengthy period in the tissue culture conditions used.

scholarly journals Cycle initiation and colony formation in culture by murine marrow cells with long-term reconstituting potential in vivo

Blood ◽  
1996 ◽  
Vol 88 (11) ◽  
pp. 4149-4158 ◽  
Author(s):  
M Trevisan ◽  
XQ Yan ◽  
NN Iscove
Keyword(s):  
Colony Formation ◽  
Interleukin 1 ◽  
Methyl Cellulose ◽  
Culture Conditions ◽  
Liquid Suspension ◽  
Almost All ◽  
Marrow Cells

Abstract This investigation was directed at separating long-term reconstituting (LTR) stem cells in normal murine marrow from hematopoietic precursors detectable in short-term assays in vitro and in vivo, and then at determining whether purified LTR cells could themselves form colonies in culture. To do so, it was first necessary to identify culture conditions that would induce their growth while preserving their long- term reconstituting capacity. Marrow was cultured with various cytokines in liquid suspension for 4 days, after which the surviving LTR activity was quantitated in a competitive in vivo assay. Activity was preserved near input levels with combined murine c-kit ligand (KL), interleukin-1 (IL-1), IL-6, and IL-11. When the cultures also included tritiated or unlabeled thymidine, LTR potential was eliminated, indicating that essentially all LTR cells were induced into cell cycle with these cytokines. To purify them, marrow was sorted on the basis of Ly6A expression and Rhodamine 123 retention. The Ly6AhiRh123ls fraction contained 85% of total recovered LTR activity but only 1% of the recovered cells measured by multilineage colony formation in spleens or in vitro. This fraction was cultured in methyl cellulose with KL, IL-1, IL-6, and IL-11 for 4 to 6 days, after which colonies were isolated and injected into mice. High levels of permanent reconstitution were achievable in sublethally irradiated W41/W41 mice after the injection of a single reconstituting unit, and limiting dilution analysis estimated the frequency of multilineage LTR at 1 in 11,200 unpurified adult marrow cells. In either lethally irradiated normal or sublethally irradiated W41/W41 mice, 1-year lymphomyeloid reconstitutions were obtained from 1 in 65 to 84 colonies of 2 to 16 dispersed cells, but not from larger colonies or those with clumped cells. The results establish that resting marrow LTR cells can be separated from almost all of the more advanced clonogenic cells that are still pluripotential, can be induced to cycle in culture by defined cytokines with preservation of their reconstituting potential, and can be manipulated and assayed efficiently at single-cell and colony levels.

Long-term passage of tachyzoites in tissue culture can attenuate virulence of Neospora caninum in vivo

Parasitology ◽  
2006 ◽  
Vol 133 (4) ◽  
pp. 421-432 ◽  
Author(s):  
P. M. BARTLEY ◽  
S. WRIGHT ◽  
J. SALES ◽  
F. CHIANINI ◽  
D. BUXTON ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Body Weight ◽  
Growth Rates ◽  
Clinical Symptoms ◽  
Neospora Caninum ◽  
High Passage ◽  
Low Passage

To determine whether prolonged in vitro passage would result in attenuation of virulence in vivo, Neospora caninum tachyzoites were passaged for different lengths of time in vitro and compared for their ability to cause disease in mice. Groups of Balb/c mice were inoculated intraperitoneally with 5×106 or 1×107 of low-passage or high-passage N. caninum tachyzoites. The mice were monitored for changes in their demeanour and body weight, and were culled when severe clinical symptoms of murine neosporosis were observed. Mice inoculated with the high-passage parasites survived longer (P<0·05), and showed fewer clinical symptoms of murine neosporosis, compared to the mice receiving the low-passage parasites. The parasite was detected in the brains of inoculated mice using immunohistochemistry and ITS1 PCR. Tissue cysts containing parasites were seen in mice inoculated with both low-passage and high-passage parasites. When the in vitro growth rates of the parasites were compared, the high-passage parasites initially multiplied more rapidly (P<0·001) than the low-passage parasites, suggesting that the high-passage parasites had become more adapted to tissue culture. These results would suggest that it is possible to attenuate the virulence of N. caninum tachyzoites in mice through prolonged in vitro passage.

scholarly journals Cycle initiation and colony formation in culture by murine marrow cells with long-term reconstituting potential in vivo

Blood ◽  
1996 ◽  
Vol 88 (11) ◽  
pp. 4149-4158 ◽  
Author(s):  
M Trevisan ◽  
XQ Yan ◽  
NN Iscove
Keyword(s):  
Colony Formation ◽  
Interleukin 1 ◽  
Methyl Cellulose ◽  
Culture Conditions ◽  
Liquid Suspension ◽  
Almost All ◽  
Marrow Cells

This investigation was directed at separating long-term reconstituting (LTR) stem cells in normal murine marrow from hematopoietic precursors detectable in short-term assays in vitro and in vivo, and then at determining whether purified LTR cells could themselves form colonies in culture. To do so, it was first necessary to identify culture conditions that would induce their growth while preserving their long- term reconstituting capacity. Marrow was cultured with various cytokines in liquid suspension for 4 days, after which the surviving LTR activity was quantitated in a competitive in vivo assay. Activity was preserved near input levels with combined murine c-kit ligand (KL), interleukin-1 (IL-1), IL-6, and IL-11. When the cultures also included tritiated or unlabeled thymidine, LTR potential was eliminated, indicating that essentially all LTR cells were induced into cell cycle with these cytokines. To purify them, marrow was sorted on the basis of Ly6A expression and Rhodamine 123 retention. The Ly6AhiRh123ls fraction contained 85% of total recovered LTR activity but only 1% of the recovered cells measured by multilineage colony formation in spleens or in vitro. This fraction was cultured in methyl cellulose with KL, IL-1, IL-6, and IL-11 for 4 to 6 days, after which colonies were isolated and injected into mice. High levels of permanent reconstitution were achievable in sublethally irradiated W41/W41 mice after the injection of a single reconstituting unit, and limiting dilution analysis estimated the frequency of multilineage LTR at 1 in 11,200 unpurified adult marrow cells. In either lethally irradiated normal or sublethally irradiated W41/W41 mice, 1-year lymphomyeloid reconstitutions were obtained from 1 in 65 to 84 colonies of 2 to 16 dispersed cells, but not from larger colonies or those with clumped cells. The results establish that resting marrow LTR cells can be separated from almost all of the more advanced clonogenic cells that are still pluripotential, can be induced to cycle in culture by defined cytokines with preservation of their reconstituting potential, and can be manipulated and assayed efficiently at single-cell and colony levels.

scholarly journals The establishment of in vivo-like tissue culture conditions in ThinCert™ tissue culture products

BioTechniques ◽  
2007 ◽  
Vol 43 (6) ◽  
pp. 812-813
Author(s):  
Sven Mühlfriedel ◽  
Günther Knebel
Keyword(s):  
Tissue Culture ◽  
Culture Conditions

The regenerative capacity of the notochordal cell: tissue constructs generated in vitro under hypoxic conditions

2009 ◽  
Vol 10 (6) ◽  
pp. 513-521 ◽  
Author(s):  
W. Mark Erwin ◽  
Facundo Las Heras ◽  
Diana Islam ◽  
Michael G. Fehlings ◽  
Robert D. Inman
Keyword(s):  
Extracellular Matrix ◽  
Tissue Culture ◽  
Sodium Alginate ◽  
Cultured Cells ◽  
3D Culture ◽  
Culture Conditions ◽  
Notochordal Cells ◽  
Collagen Ii

Object The intervertebral disc (IVD) is a highly avascular structure that is occupied by highly specialized cells (nucleus pulposus [NP] cells) that have adapted to survive within an O2 concentration of 2–5%. The object of this study was to investigate the effects of long-term hypoxic and normoxic tissue cultures of nonchondrodystrophic canine notochordal cells—cells that appear to protect the disc NP from degenerative change. Methods The authors obtained notochordal cells from nonchondrodystrophic canines according to their established methods and placed them into monolayer and 3D culture using sodium alginate globules under either hypoxic (3.5% O2) or normoxic (21% O2) conditions. Histological, immunohistochemical, scanning electron microscopy, and histomorphometric methods were used to evaluate the cells within the globules after 5 months in culture. Results Notochordal cells under in vitro hypoxic tissue culture conditions produced a highly complex, organized, 3D cellular construct that was strikingly similar to that observed in vivo. In contrast, traditional normoxic tissue culture conditions resulted in notochordal cells that failed to produce an organized matrix. Hypoxia resulted in a matrix rich in aggrecan and collagen II, whereas normoxic cultured cells did not produce any observable aggrecan or collagen II after 5 months of culture. Conclusions Hypoxia induces notochordal cells to organize a complex 3D cellular/extracellular matrix without an external scaffold other than suspension within sodium alginate. These cells produce an extracellular matrix and large construct that shares exactly the same characteristics as the in vivo condition—robust aggrecan, and type II collagen production. Normoxic tissue culture conditions, however, lead to a failure of these cells to thrive and a lack of extracellular matrix production and significantly smaller cells. The authors suggest that future studies of NP cells and, in particular, notochordal cells should utilize hypoxic tissue culture conditions to derive meaningful, biologically relevant conclusions concerning possible biological/molecular interventions.

scholarly journals ANÁLISE DOS GRÃOS DE PÓLEN DE DIFERENTES CULTIVARES DE MANONA (RICINUS COMMUNIS L., EUPHORBIACEAE): CONSERVAÇÃO E VIABILIDADE

2009 ◽  
Vol 76 (1) ◽  
pp. 115-120
Author(s):  
D.P. Vargas ◽  
S.A.M. SOUZA ◽  
S.D. Anjos e SILVA ◽  
V.L. Bobrowski
Keyword(s):  
Ricinus Communis ◽  
Ricinus Communis L

RESUMO A análise da fertilidade dos grãos de pólen de Ricinus communis (mamona) é de grande importância para programas de melhoramento, permitindo o manejo e uso adequado das coleções existentes e a criação de cultivares interessantes na produção de biodiesel. Neste trabalho, optou-se por um processo preditivo baseado no método citológico de coloração dos grãos de pólen e no método de germinação in vitro dos grãos, após tratamentos de conservação a baixas temperaturas, para a verificação da viabilidade, importante na formação das sementes, alvo dos programas de melhoramento. Utilizou-se a técnica de coloração com carmim acético 2% na análise da viabilidade dos grãos de pólen nas cultivares IAC-80, Cafelista, AL-Preta e AL-Guarany 2002, enquanto que o método de germinação in vitro após tratamento de -196° C, -80° C e -18° C, por 15 e 30 dias, foi aplicado somente na cultivar IAC-80. A viabilidade polínica foi acima de 95% em IAC-80 e Cafelista de 88,48% em AL-Preta e de 86,46% em e AL-Guarany 2002. Estas cultivares apresentaram também alto percentual de viabilidade polínica na antese, sendo possível a utilização de todas as cultivares analisadas na indução de fertilização em programas de melhoramento. Os resultados obtidos com a germinação in vitro dos grãos de IAC-80, após a criopreservação, indicaram que o percentual médio da viabilidade polínica in vivo não foi influenciado pelo período de armazenamento. Observaram-se diferenças entre os tratamentos a baixas temperaturas, porém o percentual de viabilidade dos mesmos foi baixo, sugerindo necessidade de adequação na técnica de avaliação.

scholarly journals MICROPROPOGATION AND ANALYSIS OF PHYTOCHEMICAL PROFILE OF TISSUE CULTURE GROWN PLANTAGO OVATA FORSK.

2017 ◽  
Vol 10 (4) ◽  
pp. 202 ◽  
Author(s):  
Mamta Sharma ◽  
Amita Kumari ◽  
Eshita Mahant
Keyword(s):  
Tissue Culture ◽  
Ecological Factors ◽  
Culture Method ◽  
Culture Conditions ◽  
Culture Technique ◽  
Plantago Ovata ◽  
Tissue Culture Technique ◽  
Grown Plant

Objectives : Plantago ovata is an important medicinal plant of Himalayan region greatly used in herbal dugs manufacturing. The plant is multipurpose and strictly present in the Himalaya. Plantago has many medicinal properties such as antioxidant, anti-inflammatory and hematopoiesis effects and protects the liver and is used for the treatment of cancer. The plant being medicinal possesses complex phytochemicals. The investigation of various Plantago organ (leaves, stem etc) revealed their high potential to produce a wide array of bioactive secondary metabolites. In present study the a new method of micropropagation through tissue culture  was developed for Plantago so as to meet the future demand of plant. Futher a morphological and physiochemical comparison of tissue culture grown plant was done with in vivo grown plants.Methods:  Plantago ovata was grown in -vitro through tissue culture technique using MS media and in-vivo in the nursery area of Shoolini University. In vitro culture of  Plantago ovata forsk. were managed to restrict the ecological factors and to control the culture conditions. Experimental culture parameter including germination and phytochemical constituents of Plantago ovata in vivo and in vitro conditions were observed.Results: The result revealed changes in the concentration of phytochemical constituent’s in tissue culture grown Plantago. Phytochemicals constituents (carbohydrate, tannin, chlorophyll, saponin) was reduced in tissue culture grown plant where as some phytochemicals (phenol, alkaloid, flavanoid, protein, phytosterol) increased in tissue culture grown plant than in vivo plant.  A reduction in morphological trait was found in tissue cultured plant.Conclusion: The developed tissue culture method for the micropropagation of  Plantago ovata can be used as milestone to meet the industrial need in near future.Keywords: Plantago, Tissue Culture Technique, germination, phytochemicals.

Direct Evidence for Multiple Self-Renewal Divisions of Human In Vivo Repopulating Hematopoietic Cells in Short-Term Culture

Blood ◽  
1999 ◽  
Vol 94 (7) ◽  
pp. 2161-2168 ◽  
Author(s):  
H. Glimm ◽  
C.J. Eaves
Keyword(s):  
Fetal Liver ◽  
Hematopoietic Cells ◽  
Culture Conditions ◽  
Short Term ◽  
Human Cord Blood ◽  
Self Renewal ◽  
Rare Cells

Recently, culture conditions that stimulate the proliferation of primitive hematopoietic cells defined by various phenotypic and functional endpoints in vitro have been identified. However, evidence that they support a high probability of self-renewal leading to a large net expansion in vitro of transplantable cells with lympho-myeloid repopulating ability has been more difficult to obtain. The present study was designed to investigate whether the low overall expansion of human repopulating hematopoietic cells seen in vitro reflects a selective unresponsiveness of these rare cells to the growth factors currently used to stimulate them or, alternatively, whether they do proliferate in vitro but lose engrafting potential. For this, we used a high-resolution procedure for tracking and reisolating cells as a function of their proliferation history based on the loss of cellular fluorescence after staining with (5- and 6-) carboxyfluorescein diacetate succinimidyl ester. The results show that the vast majority of long-term culture-initiating cells and in vivo lympho-myeloid competitive repopulating units present in 5-day suspension cultures initiated with CD34+ human cord blood and fetal liver cells are the progeny of cells that have divided at least once in response to stimulation by interleukin-3, interleukin-6, granulocyte colony-stimulating factor, Steel factor, and Flt3-ligand. Thus, most human repopulating cells from these two sources are stimulated to undergo multiple divisions under currently used short-term suspension culture conditions and a proportion of these retain engraftment potential.

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