Aspects Of Tissue Culture Research Articles

Primary Cardiac Fibroblast Cell Culture: Methodological Considerations for Physiologically Relevant Conditions.

Primary cardiac fibroblast (CF) tissue culture is a necessary tool for interrogating specific signaling mechanisms that dictate the phenotypic heterogeneity observed in vivo in different disease states. Traditional approaches that utilize tissue culture plastic and nutrient rich medium have been shown to induce CF activation and therefore alter CF subpopulation composition. This shift away from in vivo phenotypes complicate interpretation of results through the lens of the animal model. As the field works to identify CF diversity, these methodological flaws have begun to be addressed and more studies are focused on the dynamic interaction of CFs with their environment. This review focuses on the aspects of tissue culture that impact CF activation and therefore require consideration when designing in vitro experiments. The complexity of CF biology overlaid onto diverse model systems highlight the need for study-specific optimization and validation.

Biotechnological Approaches for Production of Artemisinin, an Anti-Malarial Drug from Artemisia annua L.

Artemisinin is an anti-malarial sesquiterpene lactone derived from Artemisia annua L. (Asteraceae family). One of the most widely used modes of treatment for malaria is an artemisinin-based combination therapy. Artemisinin and its associated compounds have a variety of pharmacological qualities that have helped achieve economic prominence in recent years. So far, research on the biosynthesis of this bioactive metabolite has revealed that it is produced in glandular trichomes and that the genes responsible for its production must be overexpressed in order to meet demand. Using biotechnological applications such as tissue culture, genetic engineering, and bioreactor-based approaches would aid in the upregulation of artemisinin yield, which is needed for the future. The current review focuses on the tissue culture aspects of propagation of A. annua and production of artemisinin from A. annua L. cell and organ cultures. The review also focuses on elicitation strategies in cell and organ cultures, as well as artemisinin biosynthesis and metabolic engineering of biosynthetic genes in Artemisia and plant model systems.

Medical Cannabis and Industrial Hemp Tissue Culture: Present Status and Future Potential.

In recent years high-THC (psychoactive) and low-THC (industrial hemp) type cannabis (Cannabis sativa L.) have gained immense attention in medical, food, and a plethora of other consumer product markets. Among the planting materials used for cultivation, tissue culture clones provide various advantages such as economies of scale, production of disease-free and true-to-type plants for reducing the risk of GMP-EuGMP level medical cannabis production, as well as the development and application of various technologies for genetic improvement. Various tissue culture methods have the potential application with cannabis for research, breeding, and novel trait development, as well as commercial mass propagation. Although tissue culture techniques for plant regeneration and micropropagation have been reported for different cannabis genotypes and explant sources, there are significant variations in the response of cultures and the morphogenic pathway. Methods for many high-yielding elite strains are still rudimentary, and protocols are not established. With a recent focus on sequencing and genomics in cannabis, genetic transformation systems are applied to medical cannabis and hemp for functional gene annotation via traditional and transient transformation methods to create novel phenotypes by gene expression modulation and to validate gene function. This review presents the current status of research focusing on different aspects of tissue culture, including micropropagation, transformation, and the regeneration of medicinal cannabis and industrial hemp transformants. Potential future tissue culture research strategies helping elite cannabis breeding and propagation are also presented.

Metabolic Labeling of Cultured Mammalian Cells for Stable Isotope-Resolved Metabolomics: Practical Aspects of Tissue Culture and Sample Extraction.

Stable isotope-resolved metabolomics (SIRM) methods are used increasingly by cancer researchers to probe metabolic pathways and identify vulnerabilities in cancer cells. Analytical and computational advances are being made constantly, but tissue culture and sample extraction procedures are often variable and not elaborated in the literature. This chapter discusses basic aspects of tissue culture practices as they relate to the use of stable isotope tracers and provides a detailed metabolic labeling and metabolite extraction procedure designed to maximize the amount of information that can be obtained from a single tracer experiment.

Effects of L-tryptophan, L-proline, and activated charcoal on plant regeneration in indica rice (Oryza sativa L.)

Production and maintenance of embryogenic calli and subsequent plant regeneration in higher frequency are important aspects of tissue culture. A rapid decrease in morphogenetic capacity with age in culture has often been observed. To overcome this problem, several methods have been suggested. We report the promotive effect of L-tryptophan, L-proline, and activated charcoal on plant regeneration in long-term callus cultures of indica rice.

Seaweed micropropagation techniques and their potentials: an overview

The seaweed industry worldwide uses 7.5–8.0 million tonnes of wet seaweeds annually with a majority of it derived from cultivated farms, as the demand for seaweed based-products exceeds the supply of seaweed raw material from natural stocks. The main advantage of cultivation is that it not only obviates overexploitation of natural populations but also facilitates the selection of germplasm with desired traits. To enhance the economic prospects of seaweed cultivation, varied practices, such as simple and cost effective cultivation methods, use of select germplasm as seed stock coupled with good farm management practices, etc., are adopted. Nevertheless, in vitro cell culture techniques have also been employed as they facilitate development and propagation of genotypes of commercial importance. There are more than 85 species of seaweeds for which tissue culture aspects have been reported. Although the initial aim of these techniques focuses mostly on genetic improvement and clonal propagation of seaweeds for mariculture, recently the scope of these techniques has been extended for use in bioprocess technology for production of high value chemicals of immense importance in the pharmaceutical and nutraceutical sectors. Recently, there has been a phenomenal interest in intensifying seaweed tissue and cell culture research to maximize the add-on value of seaweed resources. This paper deals with the status of seaweed micropropagation techniques and their applications in the context of the marine biotech industry. Further, it also provides an analysis of the problems to be resolved for removing the barriers that are impeding the true realization of potentials offered by these techniques for sustainable development and utilization of seaweed resources.

Tissue Culture Studies of Tomato (Lycopersicon esculentum)

Tomato is a major vegetable crop that has achieved tremendous popularity over the last century. It is grown in almost every country of the world. Development of protocols for in vitro selection can provide new advances for the production of stress tolerant cultivars. Techniques have been optimised for the production of haploids and somatic hybrids. Attempts have also been made to transfer the higher regenerative ability of wild varieties to cultivated tomatoes. Although, some information is available on the morphogenesis of tomato, the techniques have not been developed to a level at which they can be utilised in large-scale multiplication of commercially important cultivars. The morphogenesis response seems to be highly dependent PGRs used in the media, which is again cultivar and genotypic specific. Somatic embryogenesis in tomato is still at its infancy, and efficient procedures for large-scale production via somatic embryogenesis are yet to be developed. Genetic stability of the tissue culture raised tomato plants also needs to be addressed. The use of a combination of molecular and conventional breeding techniques could be the option for the development of cultivars resistant to biotic and abiotic stresses. This paper reviews the advances made in various aspects of tissue culture in tomato. It also discusses the issues that still need to be addressed to utilise the full potential of plant tissue culture techniques in genetic improvement and mass propagation of tomato.

Production of azadirachtin from plant tissue culture: State of the art and future prospects

With increasing awareness towards environment-friendly and non-toxic pesticide azadirachtin obtained from neem tree (Azadirachta indica) is gaining more and more importance. Its broad-spectrum activity, peculiar mode of action. eco-friendly and non-toxic action towards beneficial organisms has offered many advantages over chemical pesticides. All currently use commercial formulations based on azadirachtin contains azadirachtin extracted from seeds of naturally grown whole plants which is labour intensive process depending upon many uncontrollable geographical and climatic factors. Plant tissue culture can be a potential process for the production, offering consistent, stable and controlled supply of this bioactive compound, However the research on tissue culture aspects of production are in preliminary stage and requires culture and process optimization for the development of a commercially viable process. This review states the present status and future challenges of plant tissue culture for azadirachtin production.

Weaning and establishment of pineapple (Ananas Comosus) plantlets in compost

Plant establishment ex-vitro is one of the most difficult aspects of tissue culture (in-vitro propagation). The paper reports on establishment after treatment with two different auxins in-vitro. About ninety-nine (99) percent of the transported plantlets successfully established in compost. Spiny individuals increased with higher NAA concentrations but leaf density decreased with higher concentration of the auxin. Spinelessness, however, increased with higher IBA than its NAA counterparts, with the highest leaf number at 1 mg/L IBA. The extension/growth of the original shoot on both IBA and NAA media were not significantly different. The highest leaf expansion was recorded on the 0.5 mg/L NAA. JOURNAL OF THE GHANA SCIENCE ASSOCIATION Volume 1 Number 1, July (1998) pp. 50-54

Genetic analysis of anther and leaf disc culture in two clones of Solanum chacoense Bitt. and their reciprocal hybrids

Two diploid clones of self-incompatible Solanum chacoense Bitt. with androgenetic ability were tested for anther and leaf disc culture response together with eight of their reciprocal F1 hybrids. Large differences were found among genotypes in frequency of anther induction as well as in the phase of plant regeneration. Anthers harvested in June showed a significantly higher percentage of response (17.5%) at the induction phase than those collected in July (13.8%) or August (12.7%). The lowest induction frequency was observed in May (7.3%). By contrast, plant regeneration from induced anthers did not vary during this time. Genotypic differences were also observed in leaf disc response. The two parental clones and two of their hybrids failed to produced any shoots. Among the remaining genotypes, two had only sporadic occurrence of shoot formation, two gave an intermediate response (15% and 24% of their discs carried shoots), whereas the discs of the two remaining genotypes responded well to culture (68% and 77%). The genetic analysis performed on the reciprocal hybrids revealed that a positive significant correlation existed between anther induction and leaf disc response (Spearman's r=0.82; p=0.01). This suggests that, under our conditions, these two aspects of tissue culture might share a common system of genetic control. Estimates of broad sense heritabilities, for leaf disc culture, 83% were obtained and the number of effective factors involved in the control of tissue culture response, indicated a relatively simple genetic control. Finally, considering the potentialities opened by the use of RFLP analysis, it might be possible to find probes that are linked with genes involved in tissue culture competence.

The Eye and Inborn Errors of Metabolism

The Eye and Inborn Errors of Metabolism is another in the Birth Defects series sponsored by the National Foundation—March of Dimes. This volume records 42 papers with discussions presented at a symposium in Oxford, England, in April 1975. As one would expect with such a broad topic, the papers are diverse with respect to disease as well as approach. Many heritable metabolic disorders affecting the eye are mentioned from at least one aspect. An incomplete list of those considered in more detail includes the mucopolysaccharidoses, the mucolipidoses, the sphingolipidoses, tyrosinemia, cystinosis, Wilson disease, fucosidosis, the dyslipoproteinemias, familial LCAT deficiency, alkaptonuria, galactosemia, mannosidosis, homocystinuria, retinal degenerations, gyrate atrophy with hyperornithinemia, albinism, sickle cell retinopathy, Menkes disease, osteogenesis imperfecta, and Lowe syndrome. Many presentations are concerned predominantly with biochemistry, histochemistry, electron microscopy, or tissue culture aspects, while some are very clinically oriented. The papers are well written, richly illustrated, and superbly referenced.

Antenatal diagnosis of inborn errors of metabolism: tissue culture aspects.

The cells from 62 amniotic fluids have been cultured to the stage at which biochemical studies could have been undertaken. Although all cultures showed initial signs of cellular proliferation in only 90% of these were sufficient cells obtained for biochemical assay. If a time limit of 6 weeks was to be imposed, only 58% of the cultures could have been regarded as successful. The problems involved in culturing amniotic fluid cells for the antenatal diagnosis of inborn errors of metabolism are discussed.

Microbiological Aspects of Tissue Culture

Microbiological Aspects of Tissue Culture