scholarly journals DIFFERENTIAL EFFECT OF PURIFIED QUERCETIN AND ITS DERIVATIVES FROM IN VITRO CELL SUSPENSION CULTURES OF CAESALPINIA PULCHERRIMA SW. AGAINST SELECTED CANCER CELL LINES AND ITS MODE OF ACTION

2018 ◽  
Vol 8 (5-s) ◽  
pp. 196-208
Author(s):  
JM Aswathy ◽  
Greeshma Murukan ◽  
Bosco Lawarence ◽  
K Murugan
Keyword(s):  
Flow Cytometry ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Lines ◽  
Cell Suspension Culture ◽  
Callus Induction ◽  
Mode Of Action ◽  
Cell Suspension Cultures ◽  
Mcf 7

Tribal people use the floral extract of Caesalpinia pulcherrima to cure liver, stomach and skin prone disorders in traditional Indian medicine. This study aimed to evaluate the effect of purified quercetin and its derivatives from in vitro cell suspension cultures of C. pulcherrima Sw. against SW 480, HeLa, MCF-7 and MCF 10A cell lines and its mode of action. Standard protocol was developed for callus induction using leaf explants. Cytotoxic effect was evaluated against SW 480, HeLa, MCF-7 and MCF 10A cells by MTT assay. Apoptosis was evaluated via Hoechst analysis,  flow cytometry, mitochondrial membrane potential and caspase 3 and 9 expression. 2, 4-D (2.5 mg/l), BAP (2.5 mg/l) + kin (1 mg/ml) was effective for remarkable callus induction. Further, cell suspension culture was established.  Effect of elicitors on cell suspension culture was also carried. Sucrose, ABA and salicylic acid (SA) at different concentrations influenced cell biomass and quercetin synthesis. Cells cultured on the medium fortified with 45 g/L sucrose without ABA/SA showed the highest quercetin content (16.5 mg/g). Quercetin was purified, fractionated by HPLC-DAD and was further analyzed by NMR revealed a major fraction of quercetin (3, 5, 7, 3’, 4’-pentahydroxyflavon). Insignificant cytotoxicity was noticed in SW 480, HeLa, MCF 10A when compared to MCF-7 cell lines exposed to different concentrations of purified quercetin for 24- 48 h. Similarly, the apoptosis by nuclei staining using Hoechst 33258 revealed a concentration dependent effect on MCF 7 cells only. This was further substantiated by caspase-9 and 3 induction and mitochondrial depolarization as revealed by flow cytometry. Overall, the results showed that quercetin and its derivatives induced effective apoptosis on MCF-7 cells. Quercetin isolated from the in vitro cell suspension culture of C. pulcherrima showed significant cytotoxicity and apoptotic activity towards MCF-7 cell lines as compared to other cell lines. Keywords:  Caesalpinia pulcherrima; quercetins; suspension culture; cytotoxicity; apoptotic.

scholarly journals INDUKSI BIAK KALUS DAN BIAK SUSPENSI SEL Aquilaria malaccensis Lam.

2017 ◽  
Vol 16 (1) ◽  
pp. 1-11
Author(s):  
Aryani Leksonowati ◽  
Witjaksono Witjaksono ◽  
Diah Ratnadewi
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Callus Formation ◽  
Combined Treatment ◽  
Cell Suspension Cultures ◽  
Compact Structure ◽  
Friable Callus ◽  
Aquilaria Malaccensis

Aquilaria malaccensis Lam. is a plant species producing fragrant woody material that contains some resin. The compounds can be used as medicine and perfume. Sesquiterpenoid, one group of compounds has been found being synthesized and subsequently extracted from callus and cell suspension culture of Aquilaria species. The aim of this research was to find a method of producing friable calli and cell suspension cultures from leaves or internodes of A. malaccensis in vitro by using suitable plant growth regulators; cell suspension that will suitably serve as material to produce sesquiterpenoid afterwards. Calli were established in almost all treatments of auxin-cytokinin on both leaves and internod explants. The treatment of 10 mg/L IBA induced the highest percentage of callus coverage from leaves with a rather compact structure. The combined treatment of 1–2 mg/L 2.4-D and 0.2–0.3 mg/L BA induced friable callus formation in more than 80% of cultures with 27–32% callus coverage percentage.  The use of 2,4-D induced a better formation of cell suspension than Picloram, with maximum volume up to 7 mL. Cell suspension culture with fine and homogenous aggregate could be established in the medium supplemented with 0.5 –1 mg/L 2,4-D.

scholarly journals Potential Embryogenic Callus Induction Protocol Through Cell Suspension Culture For High Frequency Plant Regeneration Of Maspine Pineapple (Ananas comosus L.)

1970 ◽  
Vol 3 (2) ◽  
pp. 40-45
Author(s):  
M.F. Mohamad Bukhori ◽  
Norzulaani Khalid ◽  
Ch'ng Lou Ven
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Callus Induction ◽  
Embryogenic Callus ◽  
Ananas Comosus ◽  
Ms Medium ◽  
Embryogenic Callus Induction

To explore the potential for embryogenic callus induction protocol through cell suspension culture forhigh frequency plant regeneration of Maspine pineapple (Ananas comosus L.), eight different culturemedia formulation were evaluated for their effects on the induction of somatic embryos from suckerexplants. Explants were cultured on MS medium supplemented with various media concentration(NAA, Dicamba and BAP, Picloram, Kinetin and NAA, 2,4-D, TDZ, and TDZ and BAP).Embryogenic callus induction percentage, color and texture of the callus were assessed after fivemonths of culture. The optimum medium for the proliferation of in vitro shoots from sucker explantswas MS medium supplemented with 3 mg/L BAP. Meanwhile, the optimum medium for the inductionof fastest and high percentage of embryogenic callus growth from in vitro leaf-based was MS mediumsupplemented with Picloram. Results of mean comparison showed that 3 mg/L Picloram were moreeffective on explants than 10 mg/L. Results of the double staining method proved that somaticembryogenesis occurred in MS supplemented with 3 mg/L Picloram. Under microscopic observations,the globular-stage of the embryos were revealed in callus cells which is relatively suitable forsuspension cells inoculums, indicating that the tested PGR were significantly effective for somaticembryogenesis formation in this species. Most embryogenic callus from sucker explants wasyellowish-mucilaginous-wet-friable. The developed protocol potentially leads to the production ofembryogenic callus from sucker explants and plant regeneration through somatic embryogenesis.

scholarly journals TINJAUAN Penggunaan Suspensi Sel dalam Kultur In Vitro

2016 ◽  
Vol 5 (2) ◽  
pp. 84 ◽  
Author(s):  
Sri Hutami
Keyword(s):  
Secondary Metabolites ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Mass Production ◽  
Large Scale ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Cell Physiology

<p>Cell suspension culture could be defined as a<br />process that allows rapidly dividing homogenous suspension<br />of cells to grow in liquid nutrient media. There are two main<br />types of suspension cultures: (1) Batch cultures in which<br />cells are nurtured in a fixed volume of medium until growth<br />ceases and (2) Continuous cultures in which cell growth is<br />maintained by continuous replenishment of sterile nutrient<br />media. Plant cell suspension cultures are mostly used for the<br />biochemical investigation of cell physiology, growth, metabolism,<br />protoplast fusion, transformation and for large scale<br />production of seed by bioreactor and production of secondary<br />metabolites. Contamination is one of the largest problems<br />when dealing with cell cultures. Differences between<br />the products of cell suspension culture and whole plant are<br />frequently observed. These phenomena’s may be resulted<br />from lack of differentiation and organization and cell cultureinduced<br />variation. Utilization of cell suspension culture in<br />Indonesia is still limited, some of them for mass production<br />of plantation seed with bioreactor system and for production<br />of secondary metabolites. The success of this study give the<br />opportunity for mass production of seeds from other plants<br />and also production of secondary metabolites.</p>

scholarly journals Establishment of Callus Induction and Cell Suspension Cultures of Dendrathema Indicum var. Aromaticum a Scented Chrysanthemum

2017 ◽  
Vol 6 (2) ◽  
pp. 38 ◽  
Author(s):  
Liyan Jin ◽  
Yang Yang ◽  
Wenjie Gao ◽  
Mingxue Gong ◽  
Jijia Wang ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Callus Induction ◽  
Time Course ◽  
Callus Formation ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Growth Potential ◽  
Stem Segments

Dendranthema indicum var. aromaticum is an important aroma plant in genus Dendrathema, and the establishment of callus cultures and cell suspension cultures is the basement of further protoplast fusion studies, which make it possible to breed new fragrant chrysanthemum. In this study, the effects of different plant growth regulating substances in different concentrations on callus induction were investigated with stem segments, leaves, petioles as explants. The results showed that the optimal explants were lower stem segments according to the percentage of callus formation, callus hardness, growth potential and shoot differentiation. The optimal induction mediums were MS supplemented with 1.0 mg．l-12.4 D and 0.2 mg．l-1 6-BA. The cell suspension culture system was established by using the subculture calli. The results showed that the suitable inoculum size was 2g and the suitable cell suspension culture medium was MS supplemented with 0.2 mg．l-1 6-BA and 0.5 mg．l-1 2,4-D. The time course of cell growth showed that the greatest cell fresh weight appeared on day 14 and the highest cell viability on day 3.

scholarly journals Comparison of Purified Anthocyanin Isolated From In Vitro Cell Suspension Culture of Begonia malabarica, Begonia rex-cultorum ‘Baby Rainbow’ and its Antioxidant Activity

2017 ◽  
Vol 9 (08) ◽  
Author(s):  
Aswathy J. M. ◽  
Murugan K.
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Hydroxyl Radicals ◽  
Cell Suspension Culture ◽  
Callus Induction ◽  
Radical Scavenging ◽  
Leaf Explants ◽  
Reducing Power ◽  
Ms Medium

Anthocyanins are the most common flavonoid molecules of vegetables and fruits, especially berries. Human consumption of anthocyanins represents the highest among the flavonoids. Epidemiological studies have suggested that the consumption of anthocyanins lowers the risk of life style disorders like cardiovascular disease, diabetes, arthritis and cancer. Begonia malabarica Lam. of Begoniaceae, is used traditionally as anti-hypoglycemic, antimicrobial, wound healing and in the treatment of anemia. Begonia rex-cultorum ‘Baby rainbow’ an ornamental species was also substituted. Experimentation of in vitro cell suspension culture, isolation, purification of anthocyanin and its antioxidant potential are targeted in the present study. Explants such as leaves and nodes were cultured on MS medium with various phytohormones for callus induction. Leaf explants of Begonia cultured on MS medium fortified with 2, 4-D and BAP showed significant callus induction and also in terms of fresh and dry weights. Significant reddish coloured callus was achieved in cultures initiated from nodal explants in MS medium supplemented with 2, 4-D.Cell suspension cultures were also established in liquid MS medium. After 14 days of culture, cell suspension was obtained with optimal biomass accumulation. Subsequently, bioactive anthocyanin was isolated, purified and fractionated from B.malabarica andB. rex-cultorum‘Baby rainbow’ using amberlite column chromatography and LC-MS/MS analysis. The major anthocyanins eluted from Begonia speciesat 4.7-5.4 min. Tandem MS of the m/z 655.3 peak was identified as anthocyanidin Malvidin-3 –diglucoside as the major compound. The other peaks identified were (584.3) Malvidin or Peonidin, (459.2) Delphinidin + Glucose, (403.2) may be Cynanidin, (287.1) Cyanindin Aglycone and other m/z 242.3, 195.1 and 144.1 were sugar derivatives or fragments. Purified anthocyanin exhibited remarkable inhibition of linoleic acid oxidation and also a concentration dependent free-radical scavenging activities were noticed against DPPH•, hydroxyl radicals and superoxide anions. The degradation of deoxyribose by hydroxyl radicals was also inhibited via iron ion chelators, rather than by directly scavenging the radicals. The results are comparable with reducing power activity of ascorbate and catechin.

In Vitro Evaluation of Chitosan Induced Cytotoxicity against Cancerous Cell lines: MCF-7, Saos-2 and HeLa

2019 ◽  
Vol 19 ◽  
Author(s):  
Zeinab Abedian ◽  
Niloofar Jenabian ◽  
Ali Akbar Moghadamnia ◽  
Ebrahim Zabihi ◽  
Roghayeh Pourbagher ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Lines ◽  
Anticancer Agents ◽  
Fibroblast Cells ◽  
Apoptosis Induction ◽  
Cytotoxic Effects ◽  
Cancerous Cell ◽  
Significant Concentration ◽  
Mcf 7

Objective/ Background: Cancer is still the most common cause of morbidity in world and new powerful anticancer agents without severe side effects from natural sources is important. Methods: The evaluation of cytotoxicity and apoptosis induction was carried out in MCF-7,HeLa and Saos-2 as cancerous cell lines with different histological origin and human fibroblast served as control normal cell. The cells were treated with different concentrations of chitosan and the cytotoxicity was determined using MTT assay after 24, 48 and 72 h .The mode of death was evaluated by flow cytometry . Results: While both types of chitosan showed significant concentration-dependently cytotoxic effects against the three cancerous cell lines, fibroblast cells showed somehow more compatibility with chitosan. On the other hand, there were no significant differences between LMWC and HMWC cytotoxicity in all cell lines. The flow cytometry results showed the apoptosis pattern of death more in Saos-2 and HeLa while necrosis was more observable with MCF7. Also higher viability with both types of chitosan was seen in fibroblast as normal cells Conclusion: Chitosan shows anticancerous effect against 3 cancerous cell lines, while it is compatible with normal diploid fibroblast cells. Furthermore, it seems that the molecular weight of chitosan does not affect its anticancerous property.

scholarly journals Factors Affecting the Selection of Callus Cell Lines and the Preparation of the Cell Suspension Culture of Artemisia annua L.

2014 ◽  
Vol 23 (2) ◽  
pp. 157-163 ◽  
Author(s):  
Ch’ng Song Jin ◽  
Chan Lai Keng
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Lines ◽  
Cell Suspension Culture ◽  
Artemisia Annua ◽  
Leaf Explants ◽  
In Vitro Production ◽  
Callus Cell ◽  
Alternative Mode ◽  
Artemisia Annua L

Artemisinin, an important antimalarial drug against multidrug resistant strains of Plasmodium, can be produced in Artemisia annua L. Field production of artemisinin is affected by environmental condition and geographical location. In vitro production via cell suspension culture is an alternative mode and cell line selection is important to ensure sustainable production of biomass and artemisinin. Callus cell lines were derived from the leaf explants of five A. annua clones grown in two different locations in Vietnam. Thirty-four callus cell lines with consistent growth index (GI) were selected from these five clones and were categorized into fast (GI > 20), intermediate (GI 15 - 20) and slow (GI < 15) growing groups. The selected lines were found to have different morphology in term of colour and texture. The callus texture did affect the cell growth of A. annua in which the friable callus type showed faster, consistent and sustainable cell biomass production.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17507Plant Tissue Cult. & Biotech. 23(2): 157-163, 2013  (December)

scholarly journals Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Somatic Embryos ◽  
Culture Media ◽  
Ms Medium ◽  
Regenerated Plants ◽  
Male Flowers

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

scholarly journals Initiation of coconut cell suspension culture from shoot meristem derived embryogenic calli: A preliminary study

2016 ◽  
Vol 8 (1) ◽  
Author(s):  
U. Bhavyashree ◽  
K. Lakshmi Jayaraj ◽  
K. S. Muralikrishna ◽  
K. K. Sajini ◽  
M. K. Rajesh ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Fluorescent Microscopy ◽  
Cell Aggregation ◽  
Shoot Meristem ◽  
Malt Extract ◽  
Embryogenic Calli ◽  
Embryogenic Cell

<p>An attempt was made to establish highly competent embryogenic cell suspension culture in coconut, a species recalcitrant to in vitro culture. Embryogenic calli were initiated from shoot meristem explants of coconut. Y3 medium supplemented with 2.4-D (4.5 μM) and glutamine (34.2 μM) was found to be the best medium to initiate cell suspension. Growth evaluation was done by packed cell volume (PCV) and it was found that maximum growth volume of 9.9% was reached at 200 days of culture initiation. About 52% of viable cells were detected through fluorescent microscopy. Cell aggregation was noticed in Y3 medium supplemented with glutamine (34.2 μM), malt extract (100mg/l), biotin (40.9 μM) and kinetin (9.3 μM), but further progress could not be achieved. It was also observed that embryogenic calli were not of a friable type, but were associated with densely aggregated cells. Because of its hard nature, we were unsuccessful to obtain high quality cell suspension.</p>

Sign in / Sign up

Export Citation Format

Share Document