In Vitro Mass Cultivation of Cells and Tissues

Keyword(s):  
2018 ◽  
Vol 10 (1) ◽  
pp. 191-197
Author(s):  
Priyantini Widiyaningrum ◽  
Minnathul Khasanah ◽  
Dyah Rini Indriyanti

Many researchers confirmed that entomopathogenic nematodes (EPNs) of the genera Heterorhabditis and Steinernema can be cultivated in vitro using artificial media that containing animal nutrition. However, artificial media with insect component hasn’t been widely studied. This research aims to analyze the population of EPNs isolate of Semarang cultivated in mealworm powder media. Five doses of mealworm powder (Tenebrio molitor) were tested in this research, i.e: 0.5; 1.0; 1.5; 2.0 g.  Culture media using 1 g of mealworm larvae was used as control. The best treatment was further tested for its pathogenicity on Macrotermes sp. at seven levels of invective juveniles (IJs) : 0; 50; 100; 150; 200; 250; 300 IJs/mL. Each treatment was repeated five times. The EPNs population and termites mortality were analyzed using ANAVA, whereas pathogenic value was calculated using Probit analysis. The result showed EPNs population were significantly (LSD test; α> 0.05), likewise on termites mortality. The EPNs isolate of Semarang optimally at 0.5 g mealworm powder and  pathogenicity against termites based on LD50 and LD90 values at 220 JI/mL and 410 JI/mL doses, respectively. In conclusion, this result  can be an alternative to mass cultivation of EPNs, in effort of development of local bioinsecticides. The findings of this study also inform farmers that EPNs can be easily cultivated using the simple and available materials.


2016 ◽  
Vol 3 (2) ◽  
pp. 81
Author(s):  
Delvi Maretta ◽  
Dwi Pangesti Handayani ◽  
Henti Rosdayanti ◽  
Armelia Tanjung

Taro or Satoimo (Colocasia esculenta (L) Schott var antiquorum) is an alternative of non-rice food. To support saitomo mass cultivation in several regions in Indonesia, shoot multiplication and induction of satoimo microtuber through in vitro technique is amongst the stage to be undertaken. The aims of this study were to determine the effect of BAP (benzylaminopurine) and sucrose for shoot multiplication and microtuber induction of in vitro culture of satoimo. The experiment was arranged in two factors: BAP (0, 1, 2 and 3 mg/L) and sucrose (30, 60, 90 and 120 g/L). The result showed that the single effect of BAP or sucrose and interaction of both significantly increased the number of shoots. The effect of 2 mg/L BAP was more homogeneous than that of 1 and 3 mg/L BAP. Sucrose with the concentration of 30 g/L was the best concentration for shoot multiplication. The highest number of microtuber was achieved with 2 mg/L BAP + 30 g/L sucrose treatments, but tended to decrease due to increasing sucrose concentration. In 2 and 3 mg/L BAP treatments, the number of microtuber increased along with the increasing sucrose concentration.Keywords: satoimo, in vitro shoot, microtuber, benzylaminopurine, sucrose ABSTRAKSatoimo (Colocasia esculenta (L) Schott var antiquorum) merupakan bahan pangan alternatif non-beras. Untuk mendukung produksi massal satoimo di beberapa wilayah di Indonesia, multiplikasi tunas dan induksi umbi mikro secara in vitro merupakan tahapan yang harus dilakukan. Tujuan penelitian ini adalah untuk mengetahui pengaruh BAP dan sukrosa terhadap multiplikasi tunas dan induksi umbi mikro satoimo dalam kultur in vitro. Perlakuan terdiri dari 4 taraf konsentrasi BAP (0, 1, 2 dan 3 mg/L) dan 4 taraf konsentrasi sukrosa (30, 60, 90 dan 120 g/L). Hasil penelitian menunjukkan bahwa BAP dan sukrosa secara tunggal serta interaksinya berpengaruh nyata terhadap multiplikasi tunas in vitro. Pengaruh konsentrasi BAP 2 mg/L lebih homogen dibandingkan perlakuan BAP 1 dan 3 mg/L. Sukrosa 30 g/L merupakan konsentrasi terbaik untuk multiplikasi tunas. Umbi mikro terbanyak terdapat pada perlakuan BAP 1 mg/L + sukrosa 30 g/L tetapi cenderung mengalami penurunan jika konsentrasi sukrosa dinaikkan pada konsentrasi BAP tetap. Pada perlakuan BAP 2 dan 3 mg/L jumlah umbi mikro yang terbentuk cenderung meningkat seiring dengan peningkatan konsentrasi sukrosa.Kata kunci: satoimo, tunas in vitro, umbi mikro, benzilaminopurin, sukrosa


Author(s):  
Dyah Rini Indriyanti ◽  
Nur Lailatul Muharromah

<p>Entomopathogenic nematodes Entomopathogenic nematodes (EPN) of the genera Heterorhabditis and Steinernema are commercially used to control pest insect. EPN is widely cultivated through in-vivo and in vitro methods. This research aims to discover the abundance of EPN cultivated in various artificial media. Seven types of media composition were tested in this research: media A (yeast + soybean powder), media B (yeast + chicken liver), media C (yeast + dog food), media D (yolk + soybean powder), media E (yolk + chicken liver), media F (yolk + dog food), and media G (yeast + yolk + dog food). Each media was inoculated in 1.2x103JI/mL. The growth of EPN was observed weekly in 4 weeks. Results showed that EPN could be cultivated using various media; media D, E, F, and G. Highest abundance of EPN is found in the second week of media D for 28164 JI/ml.Cell harvesting is suggested to be conducted during the first and second week to obtain maximum abundance of EPN.</p><p><strong>How to Cite</strong></p><p>Indriyanti, D., &amp; Muharromah, N. (2016). Mass Cultivation of Entomopathogenic Nematode In Artificial Media.<em> Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(1), 111-118.</p>


Author(s):  
P.L. Moore

Previous freeze fracture results on the intact giant, amoeba Chaos carolinensis indicated the presence of a fibrillar arrangement of filaments within the cytoplasm. A complete interpretation of the three dimensional ultrastructure of these structures, and their possible role in amoeboid movement was not possible, since comparable results could not be obtained with conventional fixation of intact amoebae. Progress in interpreting the freeze fracture images of amoebae required a more thorough understanding of the different types of filaments present in amoebae, and of the ways in which they could be organized while remaining functional.The recent development of a calcium sensitive, demembranated, amoeboid model of Chaos carolinensis has made it possible to achieve a better understanding of such functional arrangements of amoeboid filaments. In these models the motility of demembranated cytoplasm can be controlled in vitro, and the chemical conditions necessary for contractility, and cytoplasmic streaming can be investigated. It is clear from these studies that “fibrils” exist in amoeboid models, and that they are capable of contracting along their length under conditions similar to those which cause contraction in vertebrate muscles.


Author(s):  
John J. Wolosewick ◽  
John H. D. Bryan

Early in spermiogenesis the manchette is rapidly assembled in a distal direction from the nuclear-ring-densities. The association of vesicles of smooth endoplasmic reticulum (SER) and the manchette microtubules (MTS) has been reported. In the mouse, osmophilic densities at the distal ends of the manchette are the organizing centers (MTOCS), and are associated with the SER. Rapid MT assembly and the lack of rough ER suggests that there is an existing pool of MT protein. Colcemid potentiates the reaction of vinblastine with tubulin and was used in this investigation to detect this protein.


Author(s):  
E. J. Kollar

The differentiation and maintenance of many specialized epithelial structures are dependent on the underlying connective tissue stroma and on an intact basal lamina. These requirements are especially stringent in the development and maintenance of the skin and oral mucosa. The keratinization patterns of thin or thick cornified layers as well as the appearance of specialized functional derivatives such as hair and teeth can be correlated with the specific source of stroma which supports these differentiated expressions.


Author(s):  
M. Kraemer ◽  
J. Foucrier ◽  
J. Vassy ◽  
M.T. Chalumeau

Some authors using immunofluorescent techniques had already suggested that some hepatocytes are able to synthetize several plasma proteins. In vitro studies on normal cells or on cells issued of murine hepatomas raise the same conclusion. These works could be indications of an hepatocyte functionnal non-specialization, meanwhile the authors never give direct topographic proofs suitable with this hypothesis.The use of immunoenzymatic techniques after obtention of monospecific antisera had seemed to us useful to bring forward a better knowledge of this problem. We have studied three carrier proteins (transferrin = Tf, hemopexin = Hx, albumin = Alb) operating at different levels in iron metabolism by demonstrating and localizing the adult rat hepatocytes involved in their synthesis.Immunological, histological and ultrastructural methods have been described in a previous work.


Author(s):  
Ann Chidester Van Orden ◽  
John L. Chidester ◽  
Anna C. Fraker ◽  
Pei Sung

The influence of small variations in the composition on the corrosion behavior of Co-Cr-Mo alloys has been studied using scanning electron microscopy (SEM), energy dispersive x-ray analysis (EDX), and electrochemical measurements. SEM and EDX data were correlated with data from in vitro corrosion measurements involving repassivation and also potentiostatic anodic polarization measurements. Specimens studied included the four alloys shown in Table 1. Corrosion tests were conducted in Hanks' physiological saline solution which has a pH of 7.4 and was held at a temperature of 37°C. Specimens were mechanically polished to a surface finish with 0.05 µm A1203, then exposed to the solution and anodically polarized at a rate of 0.006 v/min. All voltages were measured vs. the saturated calomel electrode (s.c.e.).. Specimens had breakdown potentials near 0.47V vs. s.c.e.


Author(s):  
M.J. Murphy ◽  
R.R. Price ◽  
J.C. Sloman

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.


Author(s):  
Tai-Te Chao ◽  
John Sullivan ◽  
Awtar Krishan

Maytansine, a novel ansa macrolide (1), has potent anti-tumor and antimitotic activity (2, 3). It blocks cell cycle traverse in mitosis with resultant accumulation of metaphase cells (4). Inhibition of brain tubulin polymerization in vitro by maytansine has also been reported (3). The C-mitotic effect of this drug is similar to that of the well known Vinca- alkaloids, vinblastine and vincristine. This study was carried out to examine the effects of maytansine on the cell cycle traverse and the fine struc- I ture of human lymphoblasts.Log-phase cultures of CCRF-CEM human lymphoblasts were exposed to maytansine concentrations from 10-6 M to 10-10 M for 18 hrs. Aliquots of cells were removed for cell cycle analysis by flow microfluorometry (FMF) (5) and also processed for transmission electron microscopy (TEM). FMF analysis of cells treated with 10-8 M maytansine showed a reduction in the number of G1 cells and a corresponding build-up of cells with G2/M DNA content.


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