Showing papers in "Biotechnology Research International in 2012"
TL;DR: Bromelain accounts for many therapeutic benefits like the treatment of angina pectoris, bronchitis, sinusitis, surgical trauma, and thrombophlebitis, debridement of wounds, and enhanced absorption of drugs, particularly antibiotics.
Abstract: Bromelain belongs to a group of protein digesting enzymes obtained commercially from the fruit or stem of pineapple. Fruit bromelain and stem bromelainare prepared differently and they contain different enzymatic composition. “Bromelain” refers usually to the “stem bromelain.” Bromelain is a mixture of different thiol endopeptidases and other components like phosphatase, glucosidase, peroxidase, cellulase, escharase, and several protease inhibitors. In vitro and in vivo studies demonstrate that bromelain exhibits various fibrinolytic, antiedematous, antithrombotic, and anti-inflammatory activities. Bromelain is considerably absorbable in the body without losing its proteolytic activity and without producing any major side effects. Bromelain accounts for many therapeutic benefits like the treatment of angina pectoris, bronchitis, sinusitis, surgical trauma, and thrombophlebitis, debridement of wounds, and enhanced absorption of drugs, particularly antibiotics. It also relieves osteoarthritis, diarrhea, and various cardiovascular disorders. Bromelain also possesses some anticancerous activities and promotes apoptotic cell death. This paper reviews the important properties and therapeutic applications of bromelain, along with the possible mode of action.
303 citations
TL;DR: The results of this study demonstrated the potential of BSG as a good substrate for enhanced remediation of hydrocarbon contaminated soil at low pollution concentration.
Abstract: Soil and surface water contamination by used lubricating oil is a common occurrence in most developing countries. This has been shown to have harmful effects on the environment and human beings at large. Bioremediation can be an alternative green technology for remediation of such hydrocarbon-contaminated soil. Bioremediation of soil contaminated with 5% and 15% (w/w) used lubricating oil and amended with 10% brewery spent grain (BSG), banana skin (BS), and spent mushroom compost (SMC) was studied for a period of 84 days, under laboratory condition. At the end of 84 days, the highest percentage of oil biodegradation (92%) was recorded in soil contaminated with 5% used lubricating oil and amended with BSG, while only 55% of oil biodegradation was recorded in soil contaminated with 15% used lubricating oil and amended with BSG. Results of first-order kinetic model to determine the rate of biodegradation of used lubricating oil revealed that soil amended with BSG recorded the highest rate of oil biodegradation (0.4361 day−1) in 5% oil pollution, while BS amended soil recorded the highest rate of oil biodegradation (0.0556 day−1) in 15% oil pollution. The results of this study demonstrated the potential of BSG as a good substrate for enhanced remediation of hydrocarbon contaminated soil at low pollution concentration.
119 citations
TL;DR: The data available on PAH degradation by fungi belonging to different ecophysiological groups (white-rot and litter-decomposing fungi) under submerged cultivation and during mycoremediation of PAH-contaminated soils are summarized.
Abstract: Polycyclic aromatic hydrocarbons (PAHs) are natural and anthropogenic aromatic hydrocarbons with two or more fused benzene rings. Because of their ubiquitous occurrence, recalcitrance, bioaccumulation potential and carcinogenic activity, PAHs are a significant environmental concern. Ligninolytic fungi, such as Phanerochaete chrysosporium, Bjerkandera adusta, and Pleurotus ostreatus, have the capacity of PAH degradation. The enzymes involved in the degradation of PAHs are ligninolytic and include lignin peroxidase, versatile peroxidase, Mn-peroxidase, and laccase. This paper summarizes the data available on PAH degradation by fungi belonging to different ecophysiological groups (white-rot and litter-decomposing fungi) under submerged cultivation and during mycoremediation of PAH-contaminated soils. The role of the ligninolytic enzymes of these fungi in PAH degradation is discussed.
95 citations
TL;DR: A detailed account of the advances made in the genetic transformation of coffee and their potential applications is provided.
Abstract: Coffee is one of the most important plantation crops, grown in about 80 countries across the world. The genus Coffea comprises approximately 100 species of which only two species, that is, Coffea arabica (commonly known as arabica coffee) and Coffea canephora (known as robusta coffee), are commercially cultivated. Genetic improvement of coffee through traditional breeding is slow due to the perennial nature of the plant. Genetic transformation has tremendous potential in developing improved coffee varieties with desired agronomic traits, which are otherwise difficult to achieve through traditional breeding. During the last twenty years, significant progress has been made in coffee biotechnology, particularly in the area of transgenic technology. This paper provides a detailed account of the advances made in the genetic transformation of coffee and their potential applications.
92 citations
TL;DR: Feedstocks such as corn stover, switchgrass, prairie cord grass, sawdust, pigeon pea grass, and cotton stalk were densified using a briquetting system to compare the physicochemical properties of briquettes made from different feedstocks.
Abstract: Densification of biomass can address handling, transportation, and storage problems and also lend itself to an automated loading and unloading of transport vehicles and storage systems. The purpose of this study is to compare the physicochemical properties of briquettes made from different feedstocks. Feedstocks such as corn stover, switchgrass, prairie cord grass, sawdust, pigeon pea grass, and cotton stalk were densified using a briquetting system. Physical characterization includes particle size distribution, geometrical mean diameter (GMD), densities (bulk and true), porosity, and glass transition temperature. The compositional analysis of control and briquettes was also performed. Statistical analyses confirmed the existence of significant differences in these physical properties and chemical composition of control and briquettes. Correlation analysis confirms the contribution of lignin to bulk density and durability. Among the feedstocks tested, cotton stalk had the highest bulk density of 964 kg/m3 which is an elevenfold increase compared to control cotton stalk. Corn stover and pigeon pea grass had the highest (96.6%) and lowest (61%) durability.
85 citations
TL;DR: In this study, the method investigated includes biological method of environmental sustainability which seeks to investigate the various biotechnological tools (biotools) in current use and those undergoing investigations for future use.
Abstract: The environment is a very important component necessary for the existence of both man and other biotic organisms. The degree of sustainability of the physical environment is an index of the survival and well-being of the entire components in it. Additionally, it is not sufficient to try disposing toxic/deleterious substances with any known method. The best method of sustaining the environment is such that returns back all the components (wastes) in a recyclable way so that the waste becomes useful and helps the biotic and abiotic relationship to maintain an aesthetic and healthy equilibrium that characterizes an ideal environment. In this study, the method investigated includes biological method of environmental sustainability which seeks to investigate the various biotechnological tools (biotools) in current use and those undergoing investigations for future use.
64 citations
TL;DR: The results suggested that levels of total phenolics, flavonoids and their FRAP indices exhibited specificity to different plants and their parts.
Abstract: The present study was designed to evaluate relative contribution of different polyphenols (total phenolics, flavonoids, flavonols) and their antioxidants activities in aqueous extracts of different parts of some plants; Argemone mexicana, Datura metel, Calotropis procera, Thevetia peruviana, and Cannabis sativa. The antioxidants (total phenolics, flavonoids, flavones) were determined by chemical methods. The antioxidant capacities of these extracts were evaluated by FRAP assay. The results demonstrated that phenolic content was maximally present in leaves of T. peruviana. This plant exhibited minimum phenolic content in its flower as compared to other plants. The flower of D. metel contained maximum phenolic content. The flavonoids were present in highest quantity in leaves of C. procera while T. peruviana flowers showed maximum flavonoid content. The fruits of C. sativa contained maximum quantity of flavonoid as compared to other plants tested. The flower extract of C. sativa possessed highest FRAP value followed by A. mexicana and fruit of C. procera. The values of ratios of different polyphenolic compounds present in plant extracts indicated that flower of D. metel contained maximum total flavonoids and minimum phenolics. These results suggested that levels of total phenolics, flavonoids and their FRAP indices exhibited specificity to different plants and their parts.
48 citations
TL;DR: The isolation and identification of chitInase-producing Bacillus from chitin-containing wastes, production of a thermostable and alkaline chit inasese, and enzyme characterization are reported.
Abstract: This paper reports the isolation and identification of chitinase-producing Bacillus from chitin-containing wastes, production of a thermostable and alkaline chitinasese, and enzyme characterization. Bacillus thuringiensis subsp. kurstaki HBK-51 was isolated from soil and was identified. Chitinase was obtained from supernatant of B. thuringiensis HBK-51 strain and showed its optimum activity at 110°C and at pH 9.0. Following 3 hours of incubation period, the enzyme showed a high level of activity at 110°C (96% remaining activity) and between pH 9.0 and 12.0 (98% remaining activity). Considering these characteristics, the enzyme was described as hyperthermophile-thermostable and highly alkaline. Two bands of the enzyme weighing 50 and 125 kDa were obtained following 12% SDS-PAGE analyses. Among the metal ions and chemicals used, Ni2+ (32%), K+ (44%), and Cu2+ (56%) increased the enzyme activity while EDTA (7%), SDS (7%), Hg2+ (11%), and ethyl-acetimidate (20%) decreased the activity of the enzyme. Bacillus thuringiensis subsp. kurstaki HBK-51 is an important strain which can be used in several biotechnological applications as a chitinase producer.
46 citations
TL;DR: In the present study, biosurfactant production by Bacillus subtilis SPB1 was effectively enhanced by response surface methodology and the predicted value is approximately 1.65 much higher than the original production determined by the conventional one-factor-at-a-time optimization method.
Abstract: Nutritional requirements can contribute considerably to the production cost and the bioprocess economics. Media optimisation using response surface methodology is one of the used methods to ameliorate the bioprocess economics. In the present study, biosurfactant production by Bacillus subtilis SPB1 was effectively enhanced by response surface methodology. A Plackett-Burman-based statistical screening procedure was adopted to determine the most important factor affecting lipopeptide production. Eleven variables are screened and results show that glucose, K2HPO4, and urea concentrations influence the most biosurfactant production. A Central Composite Design was conducted to optimize the three selected factors. Statistical analyses of the data of model fitting were done by using NemrodW. Results show a maximum predicted biosurfactant concentration of 2.93 (±0.32) g/L when using 15 g/L glucose, 6 g/L urea, and 1 g/L K2HPO4. The predicted value is approximately 1.65 much higher than the original production determined by the conventional one-factor-at-a-time optimization method.
46 citations
TL;DR: Cadmium is the most toxic heavy metal assessed in this work, followed by arsenic and lead, and two isolates of Methylobacterium were found to be tolerant to all three metals.
Abstract: Methylobacterium strains were isolated from mangrove samples collected in Bertioga, SP, Brazil, from locations either contaminated or uncontaminated by oil spills. The tolerances of the strains to different heavy metals were assessed by exposing them to different concentrations of cadmium, lead, and arsenic (0.1 mM, 0.5 mM, 1 mM, 2 mM, 4 mM, and 8 mM). Additionally, the genetic diversity of Methylobacterium spp. was determined by sequence analysis of the 16S rRNA genes. The isolates from the contaminated locations were grouped, suggesting that oil can select for microorganisms that tolerate oil components and can change the methylotrophic bacterial community. Cadmium is the most toxic heavy metal assessed in this work, followed by arsenic and lead, and two isolates of Methylobacterium were found to be tolerant to all three metals. These isolates have the potential to bioremediate mangrove environments contaminated by oil spills by immobilizing the heavy metals present in the oil.
41 citations
TL;DR: The highest antioxidant activity was observed in acidified methanol extracts of anthocyanin in dose-dependent manner and was directly related to the total anthcyanin found in red sorghum bran.
Abstract: There is increasing interest in natural food colorants like carotenoids and anthocyanins with functional properties. Red sorghum bran is known as a rich source for anthocyanins. The anthocyanin contents extracted from red sorghum bran were evaluated by biochemical analysis. Among the three solvent system used, the acidified methanol extract showed a highest anthocyanin content (4.7 mg/g of sorghum bran) followed by methanol (1.95 mg/g) and acetone (1 mg/g). Similarly, the highest total flavonoids (143 mg/g) and total phenolic contents (0.93 mg/g) were obtained in acidified methanol extracts than methanol and acetone extracts. To study the health benefits of anthocyanin from red sorghum bran, the total antioxidant activity was evaluated by biochemical and molecular methods. The highest antioxidant activity was observed in acidified methanol extracts of anthocyanin in dose-dependent manner. The antioxidant activity of the red sorghum bran was directly related to the total anthocyanin found in red sorghum bran.
TL;DR: Three strains have been found suitable for developing processes for alkaline cellulase production and may be used in the polishing and washing of denim processing of the textile industry.
Abstract: Alkaline cellulase producing fungi were isolated from soils of an undisturbed rain forest of Peru. The soil dilution plate method was used for the enumeration and isolation of fast growing cellulolytic fungi on an enriched selective medium. Eleven out of 50 different morphological colonies were finally selected by using the plate clearing assay with CMC as substrate at different pH values. All 11 strains produced cellulases in liquid culture with activities at alkaline pH values without an apparent decrease of them indicating that they are true alkaline cellulase producers. Aspergillus sp. LM-HP32, Penicillium sp. LM-HP33, and Penicillium sp. LM-HP37 were the best producers of FP cellulase (>3 U mL−1) with higher specific productivities (>30 U g−1 h−1). Three strains have been found suitable for developing processes for alkaline cellulase production. Soils from Amazonian rain forests are good sources of industrial fungi with particular characteristics. The results of the present study are of commercial and biological interest. Alkaline cellulases may be used in the polishing and washing of denim processing of the textile industry.
TL;DR: The result obtained suggested that for optimal and concomitant synthesis of the enzymes by Trichoderma viride BITRS-1001 in submerged fermentation, minimal medium supplemented with maltose and casein were the carbon and nitrogen substrates of choice.
Abstract: The paper investigates the effects of different commercial carbon and nitrogen sources on the concomitant synthesis of amylase and polygalacturonase enzymes with the aim of optimizing them for maximal enzyme production. The microorganism used in this work was the fungus Trichoderma viride BITRS-1001, which had been previously identified as a highly active producer of amylase and polygalacturonase enzymes. The results showed that the different commercial carbon and nitrogen substrate significantly affected the concomitant syntheses of amylase and polygalacturonase in culture media supplemented with the different commercial carbon and nitrogen substrates. The result obtained suggested that for optimal and concomitant synthesis of the enzymes by Trichoderma viride BITRS-1001 in submerged fermentation, minimal medium supplemented with maltose and casein were the carbon and nitrogen substrates of choice.
TL;DR: The results indicated that the co-culture of Y5 + CBS6054 was a satisfying combination for ethanol production from non-detoxified dilute-acid lignocellulosic hydrolysates, and showed a promising prospect for industrial application.
Abstract: Saccharomyces cerevisiae Y5 (CGMCC no. 2660) and Issatchenkia orientalis Y4 (CGMCC no. 2159) were combined individually with Pichia stipitis CBS6054 to establish the cocultures of Y5 + CBS6054 and Y4 + CBS6054. The coculture Y5 + CBS6054 effectively metabolized furfural and HMF and converted xylose and glucose mixture to ethanol with ethanol concentration of 16.6 g/L and ethanol yield of 0.46 g ethanol/g sugar, corresponding to 91.2% of the maximal theoretical value in synthetic medium. Accordingly, the nondetoxified dilute-acid hydrolysate was used to produce ethanol by co-culture Y5 + CBS6054. The co-culture consumed glucose along with furfural and HMF completely in 12 h, and all xylose within 96 h, resulting in a final ethanol concentration of 27.4 g/L and ethanol yield of 0.43 g ethanol/g sugar, corresponding to 85.1% of the maximal theoretical value. The results indicated that the co-culture of Y5 + CBS6054 was a satisfying combination for ethanol production from non-detoxified dilute-acid lignocellulosic hydrolysates. This co-culture showed a promising prospect for industrial application.
TL;DR: Sucrose and xylose as carbon sources and peptone and soy residues as nitrogen sources proved to be beneficial for GI production, and Plackett-Burman design was used to study the effect of different medium ingredients.
Abstract: The combination of medium ingredients has a profound influence on the metabolic pathways running in the microorganism which regulates the production of numerous metabolites. Glucose isomerase (GI), an enzyme with huge potential in the market, can isomerise glucose into fructose. GI is used widely for the production of High-Fructose Corn Syrup (HFCS). HFCS is used as a sweetener in food and pharmaceutical industries. Streptomyces are well-known producers of numerous enzymes including glucose isomerase. An array of 75 isolates was screened for the production of glucose isomerase. The isolate Streptomyces sp. SB-P1 was found to produce maximum amount of extracellular GI. Sucrose and raffinose among pure carbon sources and corn cob and wheat husk among crude agro residues were found to yield high enzyme titers. Potassium nitrate among pure nitrogen sources and soy residues among crude sources gave maximum production. Quantitative effect of carbon, nitrogen, and inducer on GI was also determined. Plackett-Burman design was used to study the effect of different medium ingredients. Sucrose and xylose as carbon sources and peptone and soy residues as nitrogen sources proved to be beneficial for GI production.
TL;DR: Paecilomyces lilacinus (Thom) Samson LPS 876, a locally isolated fungal strain, was grown on minimal mineral medium containing “hair waste,” a residue from the hair-saving unhairing process, and produced a protease with keratinolytic activity that could effectively remove blood stains and showed high stability toward surfactants, bleaching agents, and solvents.
Abstract: Paecilomyces lilacinus (Thom) Samson LPS 876, a locally isolated fungal strain, was grown on minimal mineral medium containing "hair waste," a residue from the hair-saving unhairing process, and produced a protease with keratinolytic activity. This enzyme was biochemically characterized. The optimum reaction conditions, determined with a response surface methodology, were 60°C and pH 6.0. It was remarkably stable in a wide range of pHs and temperatures. Addition of Ca(2+), Mg(2+), or sorbitol was found to be effective in increasing thermal stability of the protease. PMSF and Hg(2+) inhibited the proteolytic activity indicating the presence of a thiol-dependent serine protease. It showed high stability toward surfactants, bleaching agents, and solvents. It was also compatible with commercial detergents (7 mg/mL) such as Ariel, Skip, Drive, and Ace, retaining more than 70% of its proteolytic activity in all detergents after 1 h of incubation at 40°C. Wash performance analysis revealed that this protease could effectively remove blood stains. From these properties, this enzyme may be considered as a potential candidate for future use in biotechnological processes, as well as in the formulation of laundry detergents.
TL;DR: Activity analysis on several uronic acids and their derivatives suggests that the enzyme recognized only esters of 4-O-methyl-D-glucuronic acids derivatives, even with a 4-nitrophenyl aglycon but did not hydrolyze the ester of D-galacturonic acid.
Abstract: The gene encoding Schizophyllum commune glucuronoyl esterase was identified in the scaffold 17 of the genome, containing two introns of 50 bp and 48 bp, with a transcript sequence of 1179 bp. The gene was synthesized and cloned into Pichia pastoris expression vector pGAPZα to achieve constitutive expression and secretion of the recombinant enzyme in soluble active form. The purified protein was 53 kD with glycosylation and had an acidic pI of 3.7. Activity analysis on several uronic acids and their derivatives suggests that the enzyme recognized only esters of 4-O-methyl-D-glucuronic acid derivatives, even with a 4-nitrophenyl aglycon but did not hydrolyze the ester of D-galacturonic acid. The kinetic values were 0.25 mM, 16.3 μM·min−1, and 9.27 s−1 with 4-nitrophenyl 2-O-(methyl 4-O-methyl-α-D-glucopyranosyluronate)-β-D-xylopyranoside as the substrate.
TL;DR: Investigation of myostatin gene polymorphism and its association with yearling weight records in Zel sheep using PCR-RFLP and PCR-SSCP methods found that this locus was not in Hardy-Weinberg equilibrium (P < 0.05), and there was no significant effect of mystatin gene on yearling weights.
Abstract: The aim of present study was to investigate myostatin gene polymorphism and its association with yearling weight records in Zel sheep using PCR-RFLP and PCR-SSCP methods. Blood samples were collected from 200 Zel sheep, randomly, and DNA was extracted using modified salting out method. Polymerase chain reaction was carried out to amplify 337, 222, and 311 bp fragments, respectively, comprising a part of exon 3, intron 1, and intron 2 of myostatin gene. In addition, exon 3 was digested by HaeIII enzyme under RFLP method, and introns 1 and 2 were studied using SSCP. Under RFLP method, all samples showed mm genotype. Under SSCP method, intron 1 was also monomorph but intron 2 was polymorph (AA, AB, and BB). The allelic frequencies for A and B were 75.5 and 24.5%, respectively. This locus was not in Hardy-Weinberg equilibrium (), and there was no significant effect of myostatin gene on yearling weights.
TL;DR: The recombinant E. cloacae strain could be improved further by increasing its ethanol tolerance capability through media optimization and also by combining multigene cellulase expression for enhancing ethanol production from various types of lignocellulosic biomass so that it can be used for industrial level ethanol production.
Abstract: The ethanol fermenting genes such as pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adh II) were cloned from Zymomonas mobilis and transformed into three different cellulolytic bacteria, namely Enterobacter cloacae JV, Proteus mirabilis JV and Erwinia chrysanthemi and their cellulosic ethanol production capability was studied. Recombinant E. cloacae JV was found to produce 4.5% and 3.5% (v/v) ethanol, respectively, when CMC and 4% NaOH pretreated bagasse were used as substrates, whereas recombinant P. mirabilis and E. chrysanthemi with the same substrates could only produce 4%, 3.5%, 1%, and 1.5 % of ethanol, respectively. The recombinant E. cloacae strain produced twofold higher percentage of ethanol than the wild type. The recombinant E. cloacae strain could be improved further by increasing its ethanol tolerance capability through media optimization and also by combining multigene cellulase expression for enhancing ethanol production from various types of lignocellulosic biomass so that it can be used for industrial level ethanol production.
TL;DR: A novel simplified method for screening and isolation of PUFA-producing bacteria by direct visualization using the H2O2-plate assay and shows drastically reduction in the number of samples thus saves the time, effort, and cost of screening and isolating strains of bacterial PUFAs producers.
Abstract: Bacterial production of polyunsaturated fatty acids (PUFAs) is a potential biotechnological approach for production of valuable nutraceuticals. Reliable method for screening of number of strains within short period of time is great need. Here, we report a novel simplified method for screening and isolation of PUFA-producing bacteria by direct visualization using the H(2)O(2)-plate assay. The oxidative stability of PUFAs in growing bacteria towards added H(2)O(2) is a distinguishing characteristic between the PUFAs producers (no zone of inhibition) and non-PUFAs producers (zone of inhibition) by direct visualization. The confirmation of assay results was performed by injecting fatty acid methyl esters (FAMEs) produced by selected marine bacteria to Gas Chromatography-Mass Spectrometry (GCMS). To date, this assay is the most effective, inexpensive, and specific method for bacteria producing PUFAs and shows drastically reduction in the number of samples thus saves the time, effort, and cost of screening and isolating strains of bacterial PUFAs producers.
TL;DR: The Kingtai-TDZ showed a high potency and suitability for use in M. volkensii tissue culture, and the effect of TDZ on embryogenicity was significant over certain ranges of concentrations.
Abstract: Melia volkensii Gurke is a drought-tolerant tree native to East Africa’s arid and semiarid lands (ASALs), with vast but underutilized potential for agroforestry and sustainable livelihoods in the ASALs. Its cultivation is limited by difficulties in propagation via conventional means. Full exploitation of the ability of thidiazuron (TDZ) to elicit regeneration in plant tissue cultures, as sole plant growth regulator (PGR), is hampered by high costs. This study tested the effectiveness of a low-cost agrochemical TDZ for in vitro propagation of M. volkensii. Zygotic embryos from mature seeds were cultured on Gamborg’s B5 medium containing 0 to 4 mg/L of agrochemical TDZ from Kingtai Chemicals Co.,Ltd., China. Callus induction frequency was 96.67 to 100%. Significantly large callus fresh mass was produced at 0.05 mg/L TDZ concentration (ANOVA,
TL;DR: A micropropagation protocol was developed which may assist in the safeguarding and augmentation of dwindling natural populations of Clinopodium odorum (Griseb.) Harley, a critically and endangered medicinal plant, with a view to develope longer-term strategies for the transfer and reintroduction into natural habitats.
Abstract: A micropropagation protocol was developed which may assist in the safeguarding and augmentation of dwindling natural populations of Clinopodium odorum (Griseb.) Harley, a critically and endangered medicinal plant. Factors affecting culture initiation bud sprouting and growth, rooting, and acclimatization were studied, using nodal segments of in vitro germinated seedling as primary explants on six media supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) (0.5–1.5 and 2-Naphthalene acetic acid (NAA) (0.5–1.5). Best results for culture initiation with sustainable multiplication rates (100%) were obtained on WP medium without any growth regulator. WP with the addition of 0.5 : 1 or 0.5 : 1.5) of BAP and NAA promoted a higher elongation; however, the optimum number of nodes were obtained in plantlets grown on 1/2 MS with the addition of 1 : 1.5 of BAP and NAA. Culture of sectioned individual nodes transferred to the media with different rates of BAP and NAA 1/2 MS-9 (1.5 : 1.5), SH-8 (1.5 : 1.0), and 1/2 B5-4 (1.0 : 0.5) media resulted in no proliferated shoots. The in vitro plants were successfully acclimatized garden soil and sand (2 : 1) in the greenhouse, with over 90% survival rate. The in vitro-grown plants could be transferred to ex vitro conditions and the efficacy in supporting ex vitro growth was assessed, with a view to develope longer-term strategies for the transfer and reintroduction into natural habitats.
TL;DR: It is indicated that a myelomonocytoid population from pancreatic explant outgrowths has mesenchymal differentiation potential and is in line with recent data onmonocyte-derivedmesenchymic progenitors (MOMPs).
Abstract: Progenitor cells can be obtained by outgrowth from tissue explants during primary ex vivo tissue culture. We have isolated and characterized cells outgrown from neonatal mouse pancreatic explants. A relatively uniform population of cells showing a distinctive morphology emerged over time in culture. This population expressed monocyte/macrophage and hematopoietic markers (CD11b+ and CD45+), and some stromal-related markers (CD44+ and CD29+), but not mesenchymal stem cell (MSC)-defining markers (CD90− and CD105−) nor endothelial (CD31−) or stem cell-associated markers (CD133− and stem cell antigen-1; Sca-1−). Cells could be maintained in culture as a plastic-adherent monolayer in culture medium (MesenCult MSC) for more than 1 year. Cells spontaneously formed sphere clusters “pancreatospheres” which, however, were nonclonal. When cultured in appropriate media, cells differentiated into multiple mesenchymal lineages (fat, cartilage, and bone). Positive dithizone staining suggested that a subset of cells differentiated into insulin-producing cells. However, further studies are needed to characterize the endocrine potential of these cells. These findings indicate that a myelomonocytoid population from pancreatic explant outgrowths has mesenchymal differentiation potential. These results are in line with recent data onmonocyte-derivedmesenchymal progenitors (MOMPs).