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.

STUDIES ON OESTROGENS IN CATTLE

1960 ◽  
Vol XXXIII (II) ◽  
pp. 277-286 ◽  
Author(s):  
Weiert Velle ◽  
Stian Erichsen
Keyword(s):  
Tissue Culture ◽  
Living Cells ◽  
Culture Technique ◽  
Kidney Cells ◽  
Chemical Methods ◽  
Bovine Kidney ◽  
Tissue Culture Technique

ABSTRACT A review is given of previous in vitro investigations on oestrogen metabolism. In the present investigation use has been made of the tissue culture technique, whereby possible blood or serum effects on oestrogen transformations could be excluded. The conversion products were characterized by chemical methods. In the presence of bovine kidney cells grown on a medium of known composition, the following conversions were recorded: Oestrone to oestradiol-17β, oestradiol-17β to oestrone, oestradiol-17α to oestrone. Control incubations of the hormones with medium only showed that the transformations must be due to the presence of the living cells. The rate of conversion to oestrone was markedly higher for oestradiol-17β than for oestradiol-17α. As previous in vivo experiments have shown oestradiol-17α to be an important end product in the bovine, following injections of both oestrone and oestradiol-17β, the free interconvertibility of oestrone and oestradiol-17β demonstrated in the present investigation becomes significant. The findings are discussed in relation to recent observations on hormoneenzyme interrelations.

scholarly journals Modulating the Substrate Stiffness to Manipulate Differentiation of Resident Liver Stem Cells and to Improve the Differentiation State of Hepatocytes

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Angela Maria Cozzolino ◽  
Valeria Noce ◽  
Cecilia Battistelli ◽  
Alessandra Marchetti ◽  
Germana Grassi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Method ◽  
Cell Types ◽  
Culture Conditions ◽  
Precursor Cells ◽  
Substrate Stiffness ◽  
Growth And Survival ◽  
Liver Stem Cells

In many cell types, several cellular processes, such as differentiation of stem/precursor cells, maintenance of differentiated phenotype, motility, adhesion, growth, and survival, strictly depend on the stiffness of extracellular matrix that,in vivo, characterizes their correspondent organ and tissue. In the liver, the stromal rigidity is essential to obtain the correct organ physiology whereas any alteration causes liver cell dysfunctions. The rigidity of the substrate is an element no longer negligible for the cultivation of several cell types, so that many data so far obtained, where cells have been cultured on plastic, could be revised. Regarding liver cells, standard culture conditions lead to the dedifferentiation of primary hepatocytes, transdifferentiation of stellate cells into myofibroblasts, and loss of fenestration of sinusoidal endothelium. Furthermore, standard cultivation of liver stem/precursor cells impedes an efficient execution of the epithelial/hepatocyte differentiation program, leading to the expansion of a cell population expressing only partially liver functions and products. Overcoming these limitations is mandatory for any approach of liver tissue engineering. Here we propose cell lines asin vitromodels of liver stem cells and hepatocytes and an innovative culture method that takes into account the substrate stiffness to obtain, respectively, a rapid and efficient differentiation process and the maintenance of the fully differentiated phenotype.

scholarly journals In Vitro Effect of Erythropoietin on the Spleen of The Polycythemic Mouse

Blood ◽  
1967 ◽  
Vol 29 (6) ◽  
pp. 852-858
Author(s):  
YASUSADA MIURA ◽  
FUMIMARO TAKAKU ◽  
KIKU NAKAO
Keyword(s):  
Tissue Culture ◽  
Culture Method ◽  
Stem Cell Differentiation ◽  
Mouse Spleen ◽  
In Vitro Method ◽  
Heme Synthesis ◽  
Tissue Culture Method ◽  
Vitro Effect

Abstract 1. An in vitro method to observe radiosensitivity of stem cells was developed in the present study. In vivo and in vitro effect of 60Co irradiation on the erythropoietin-induced stem cell differentiation into erythroblasts was observed, using a tissue culture method of polycythemic mouse spleen. Response to erythropoietin was demonstrated by an appearance of heme synthesis and erythroblasts in spleen fragments. 2. A significant correlation between the rate of appearance of erythroblasts and heme synthesis of the spleen fragments was observed. 3. After irradiation, marked impairment of both heme synthesis and production of erythroblasts was observed, yielding D37 values in the vicinity of 70 r in vivo and 120 r in vitro irradiation, respectively. 4. Marked recovery of erythropoietin-induced heme synthesis in the polycythemic mouse spleen was observed 9 days after 300 r irradiation, with an "overshooting" phenomenon on the 12th day.

scholarly journals 3D Hepatic Organoid-Based Advancements in LIVER Tissue Engineering

2021 ◽  
Vol 8 (11) ◽  
pp. 185
Author(s):  
Amit Panwar ◽  
Prativa Das ◽  
Lay Poh Tan
Keyword(s):  
Tissue Engineering ◽  
Self Assembly ◽  
Liver Tissue ◽  
3D Culture ◽  
Culture Method ◽  
Culture Conditions ◽  
Phenotypic Properties ◽  
Liver Tissue Engineering

Liver-associated diseases and tissue engineering approaches based on in vitro culture of functional Primary human hepatocytes (PHH) had been restricted by the rapid de-differentiation in 2D culture conditions which restricted their usability. It was proven that cells growing in 3D format can better mimic the in vivo microenvironment, and thus help in maintaining metabolic activity, phenotypic properties, and longevity of the in vitro cultures. Again, the culture method and type of cell population are also recognized as important parameters for functional maintenance of primary hepatocytes. Hepatic organoids formed by self-assembly of hepatic cells are microtissues, and were able to show long-term in vitro maintenance of hepato-specific characteristics. Thus, hepatic organoids were recognized as an effective tool for screening potential cures and modeling liver diseases effectively. The current review summarizes the importance of 3D hepatic organoid culture over other conventional 2D and 3D culture models and its applicability in Liver tissue engineering.

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 Perbanyakan tanaman kina Cinchona ledgeriana Moens. dan C. succirubra Pavon melalui penggandaan tunas aksiler Propagation of cinchona plant Cinchona ledgeriana Moens. and C. succirubra Pavon through axillary buds multiplication

2016 ◽  
Vol 72 (1) ◽  
Author(s):  
. JOKO-SANTOSO ◽  
Nurita TORUAN-MATHIUS ◽  
U SASTRAPRAWIRA ◽  
G SURYATMANA ◽  
D SAODAH
Keyword(s):  
Tissue Culture ◽  
Rice Husk ◽  
Shoot Multiplication ◽  
Culture Technique ◽  
Axillary Buds ◽  
Multiplication Rate ◽  
Plant Materials ◽  
Tissue Culture Technique ◽  
Cinchona Ledgeriana

SummaryCinchona ledgeriana (Ledger) and C. succirubra (Succi) were industrial commodities which their barks of the trunk  contain alkaloid   used as raw materials in pharmaceutical, food, drug and beverages and chemical industries. The problem  faced in conventional plant propagation are. incompatibilities, high numbers of death caused by transportation, limited numbers and time consume in  plant materials  production. These problems may be  overcome by axillary buds multiplication.  The aim of the experiment were to find out propagation technology of Ledger and Succi by  tissue culture technique.  Experiments were conducted in three consecutive steps, viz the effect of (i) BAP on multiplication and growth of axillary’s bud of Ledger and Succi in vitro culture, (ii) IBA on root initiation and growth, (iii)  growth medium on the growth of plantlets in  acclimatization.The design of the experiments were Complete Randomized Design with 15 (i & ii) and four (iii) replications. The treatments were (i) 0,1,2,3,4, dan 5 mg/L BAP, (ii) 0.0; 0.5; 1.0; 1.5; 2.0; dan 2.5 mg/L IBA, and (iii) mixture of soil and rice husk charcoal (1:1), mixture of soil and compost (1:1),  mixture of soil, rice husk charcoal, and  compost  (1:1:1).  Parameters measured in the experiments were (i) the initiation of buds multiplication rate twice at axillary buds at subculture.  (ii) initiation  and  roots vigor. (iii) numbers of survived  plants and plants vigor. The explant source used derived from two-month old axillary buds cultured in Murashige and Skoog (MS) medium without growth regulator. Results of the experiment showed  that the best shoot multiplication of Ledger  and Succi  was obtained from the application of 3 mg/L BAP, with buds multipli-cation rate 7 buds/explant/month for Ledger, and 3-4 buds/explants/month for succi. The best root initation and root growth were found from the application of 2 mg/L IBA. The highest percentage of survived plantlets (100%) in acclimatization was obtained from mixture of soil and rice husk charcoal (1:1) medium.  Therefore it is  concluded that tissue culture technique could be used for planlet  mass propagation    of  elite C. Ledgeriana and C. Succirubra through axillary bud multiplication.Ringkasan Tanaman kina Cinchona ledgeriana (Ledger) dan C. succirubra (Succi)  merupakan tanaman industri yang mengandung alkaloid di dalam kulit batangnya dan berguna dalam bidang industri farmasi, makanan, minuman dan kimia. Kendala yang dihadapi dalam perbanyakan tanaman kina secara konvensional dengan sistem sambung  adalah inkompatibilitas,    kematian akibat pengangkutan cukup tinggi, jumlah bahan tanam yang diproduksi sangat terbatas dan waktu penyediaan yang cukup lama. Masalah tersebut dapat diatasi dengan menggunakan teknik kultur jaringan. Penelitian ini bertujuan untuk men-dapatkan teknologi perbanyakan tanaman kina Ledger dan Succi dengan teknik kultur jaringan. Penelitian terdiri atas (i) pengaruh BAP terhadap inisiasi dan penggandaan  tunas aksilar, (ii) pengaruh IBA terhadap inisiasi serta pertum-buhan akar planlet,   dan (iii) pengaruh beberapa medium terhadap pertumbuhan planlet dalam aklimatisasi. Percobaan menggunakan Rancangan Acak Lengkap, masing-masing diulang 15 (i & ii)  dan (iii) empat kali. Peubah yang diukur untuk percobaan (i) adalah waktu inisiasi tunas dan laju penggandaan tunas aksiler pada dua  kali  subkulur. (ii)  Waktu  inisiasi  dan vigor akar. (iii) Jumlah tanaman yang bertahan hidup setelah aklimatisasi, serta vigor tanaman. Sumber eksplan yang digunakan adalah tunas aksilar dari kecambah terpilih berumur dua bulan yang dikulturkan dalam medium Murashige dan Skoog tanpa zat pengatur tumbuh. Perlakuan untuk percobaan (i) adalah 0,0; 1,0; 2,0; 3,0; 4,0 dan  5,0 mg/L BAP, (ii) adalah 0,0; 0,5; 1,0; 1,5; 2,0; dan 2,5 mg/L IBA, sedang (iii) adalah medium tanam tanah, tanah : arang sekam (1:1),  tanah : kompos (1:1), tanah : arang sekam : kompos (1:1:1). Hasil yang diperoleh menunjukkan bahwa konsentrasi BAP terbaik untuk inisiasi dan penggandaan tunas tanaman kina Ledger dan Succi adalah 3 mg/L BAP, dengan laju penggandaan tujuh tunas/eksplan/bulan untuk Ledger dan 3-4 tunas/eksplan/bulan untuk Succi. Sedang untuk perakaran diperoleh dari medium MS dengan penambahan 2 mg/L IBA. Persentase tertinggi planlet (100%) yang mampu bertahan hidup pada aklimatisasi diperoleh dari medium campuran tanah : arang sekam (1:1). Berdasarkan hasil tersebut di atas dapat disimpulkan bahwa perbanyakan tanaman kina secara in vitro untuk menghasilkan bibit bermutu dapat dilakukan melalui teknik penggandaan tunas aksiler

scholarly journals Enrichment of Melanoma Stem-Like Cells via Sphere Assays

2021 ◽  
pp. 185-199
Author(s):  
Nabanita Mukherjee ◽  
Karoline A. Lambert ◽  
David A. Norris ◽  
Yiqun G. Shellman
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Stem Cell ◽  
Three Dimensional ◽  
Culture Conditions ◽  
Low Density ◽  
In Vitro Techniques ◽  
Sphere Culture

AbstractSphere assays are widely used in vitro techniques to enrich and evaluate the stem-like cell behavior of both normal and cancer cells. Utilizing three-dimensional in vitro sphere culture conditions provide a better representation of tumor growth in vivo than the more common monolayer cultures. We describe how to perform primary and secondary sphere assays, used for the enrichment and self-renewability studies of melanoma/melanocyte stem-like cells. Spheres are generated by growing melanoma cells at low density in nonadherent conditions with stem cell media. We provide protocols for preparing inexpensive and versatile polyHEMA-coated plates, setting up primary and secondary sphere assays in almost any tissue culture format and quantification methods using standard inverted microscopy. Our protocol is easily adaptable to laboratories with basic cell culture capabilities, without the need for expensive fluidic instruments.

scholarly journals Application of Horticultural and Tissue Culture Methods for Ex Situ Conservation of Endangered Primula farinosa L.

2020 ◽  
Vol 89 (1) ◽  
Author(s):  
Ewa Sitek ◽  
Barbara Nowak ◽  
Michał Fecowicz ◽  
Zbigniew Gajewski ◽  
Piotr Dańda ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Culture Conditions ◽  
Ex Situ ◽  
In Vitro Cultivation ◽  
Culture Methods ◽  
Transient Nature ◽  
Micropropagated Plants ◽  
The Difference

Our study aimed at active conservation of the last location of <em>Primula farinosa</em>, an endangered species in Poland, and assessed reproduction by seeds and plant propagation on sterile media in tissue culture conditions. We identified gibberellic acid (GA<sub data-id="subscript-1">3</sub>) as the key factor stimulating germination of <em>P. farinosa</em> seeds. Growing juvenile plants under controlled temperature of 18/16 °C day/night yielded good quality plant material without mycorrhization. In tissue culture, the most favorable medium for shoot propagation was MS supplemented with the lowest tested concentration of indole-3-butyric acid (IBA; 0.05 mg dm<sup data-id="superscript-1">−3</sup>) and 6-benzyl-aminopurine (BAP; 0.1 mg dm<sup data-id="superscript-2">−3</sup>). The rooting ability of shoots was high and comparable for all auxins used. 2C DNA content of seed-derived and micropropagated plants did not indicate any change in the ploidy level during in vitro cultivation. Plants derived from seeds and tissue cultures were compared in a 2-year study. Of all the characteristics compared, only the number of flowers per inflorescence was lower for micropropagated plants when compared with the seed-origin plants in the first year of observation. The difference was of transient nature and was not observed in the second year of the study. Effective protocols for in vivo and in vitro propagation of <em>P. farinosa</em> were developed, which can be used in practical species protection.

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