scholarly journals MICROPROPAGATION OF CYMBIDIUM SPP. BY SOMATIC EMBRYOGENESIS TECHNIQUE

2018 ◽  
Vol 6 ◽  
pp. 1178-1184
Author(s):  
Minh Van Tran
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Cell ◽  
Activated Charcoal ◽  
Multiple Shoots ◽  
Embryonic Cell ◽  
Cell Suspensions ◽  
Set Up ◽  
Planting Materials ◽  
In Vitro Embryogenesis

Cymbidium spp. was regularly produced through micropropagation by protocorm like bodies (PLBs) and multiple shoots; micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation to resolve the above difficulties. The method was to use PLBs as planting materials to produce somatic callus cell and embryos. Results were followed as: PLBs were cut into slices and placed on the medium for callus initiated and used as materials for embryo formation study. A fresh weight of callus were used for the experiment of about 100 mg. The medium for the initiation of embryonic callus was composed of: MS + BA (0.1 mg/l) + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1mg/l) or 2.4D (1 mg/l) and was proliferated on the medium MS + pepton (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) or 2.4D (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium consisting of MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) + BA (0.1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with BA (1 mg/l). Embryonic cell suspensions were plating and regeneration on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (0.1 mg/l) + BA (1 mg/l). Micropropagation of Cymbidium spp. via embryogenesis technique was set up to produce 3,800 plantlets per one liter of somatic embryogenesis suspension.

scholarly journals MICROPROPAGATION OF PHALAENOPSIS SPP. BY SOMATIC EMBRYOGENESIS TECHNIQUE

2018 ◽  
Vol 6 ◽  
pp. 1185-1191
Author(s):  
Minh Van Tran
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Cell ◽  
Embryonic Cell ◽  
Cell Suspensions ◽  
Ms Medium ◽  
Callus Initiation ◽  
Set Up ◽  
Planting Materials ◽  
In Vitro Embryogenesis

Phalaenopsis spp. was regularly produced through micropropagation by protocorm like bodies (PLBs); micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation. The method involved using protocorm like bodies as planting materials. PLBs were cut into slices and placed on the medium for callus initiation. The callus was initiated on the medium MS + BA (0.1 mg/l) supplemented with NAA (1 mg/l) or 2,4D (1 mg/l) and was proliferated on the medium MS + BA (0.1 mg/l) supplemented with NAA (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium MS + BA (0.1 mg/l) supplemented with NAA (0.5, 1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the MS medium supplemented with NAA (0.1 mg/l) + BA (0.5 mg/l). Embryonic cell suspensions were plated and regenerated on the medium: 1/2MS supplemented with NAA (0.1 mg/l) + BA (0.5 mg/l). Micropropagation of Phalaenopsis sp. via the embryogenesis technique was set up to produce 5,800 plantlets per one liter of somatic embryogenesis suspension.

scholarly journals Self-organising human gonads generated by a Matrigel-based gradient system

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Elizabeth Oliver ◽  
João Pedro Alves-Lopes ◽  
Femke Harteveld ◽  
Rod T. Mitchell ◽  
Elisabet Åkesson ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Germ Cell ◽  
Somatic Cell ◽  
Three Dimensional ◽  
Cell Suspensions ◽  
Testicular Development ◽  
Female Gonad ◽  
Gradient System ◽  
Three Dimensional Culture

Abstract Background Advances in three-dimensional culture technologies have led to progression in systems used to model the gonadal microenvironment in vitro. Despite demonstrating basic functionality, tissue organisation is often limited. We have previously detailed a three-dimensional culture model termed the three-layer gradient system to generate rat testicular organoids in vitro. Here we extend the model to human first-trimester embryonic gonadal tissue. Results Testicular cell suspensions reorganised into testis-like organoids with distinct seminiferous-like cords situated within an interstitial environment after 7 days. In contrast, tissue reorganisation failed to occur when mesonephros, which promotes testicular development in vivo, was included in the tissue digest. Organoids generated from dissociated female gonad cell suspensions formed loosely organised cords after 7 days. In addition to displaying testis-specific architecture, testis-like organoids demonstrated evidence of somatic cell differentiation. Within the 3-LGS, we observed the onset of AMH expression in the cytoplasm of SOX9-positive Sertoli cells within reorganised testicular cords. Leydig cell differentiation and onset of steroidogenic capacity was also revealed in the 3-LGS through the expression of key steroidogenic enzymes StAR and CYP17A1 within the interstitial compartment. While the 3-LGS generates a somatic cell environment capable of supporting germ cell survival in ovarian organoids germ cell loss was observed in testicular organoids. Conclusion The 3-LGS can be used to generate organised whole gonadal organoids within 7 days. The 3-LGS brings a new opportunity to explore gonadal organogenesis and contributes to the development of more complex in vitro models in the field of developmental and regenerative medicine.

Acceleration of Dabigatran Elimination by Activated Charcoal Perfusion and Hemodialysis in a Pig Model.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2272-2272 ◽  
Author(s):  
Jessica Lange ◽  
Christian Thiel ◽  
Karolin Thiel ◽  
Wilfried Klingert ◽  
Kathrin Klingert ◽  
...  
Keyword(s):  
Steady State ◽  
Activated Charcoal ◽  
Plasma Levels ◽  
Binding Capacity ◽  
Flow Rates ◽  
Dialysate Flow ◽  
Maximum Binding Capacity ◽  
Set Up

Abstract Abstract 2272 Background and Aim: Dialysis based approaches can provide rapid removal of dabigatran in cases of emergency due to its low protein binding of ∼35%. However the in vitro properties of these filtration devices have not yet been characterized in detail. This study in the porcine system (both in vitro and in vivo) was performed to evaluate dabigatran elimination by hemodialysis and activated charcoal perfusion as compared to normal renal elimination. Methods: Porcine blood (5L) was supplemented with 1000 ng/mL dabigatran and circulated through an circuit of tubing allowing attached to an activated charcoal filter (Absorba 300 C, Gambro). Further supplementation of dabigatran allowed the determination of maximum binding capacity of the filter. A similar set up was used to also test dialysis (Polyflux 140H, Gambro) and determine the dependence of the flow rate on dabigatran removal. Dialysate flow rates were increased up to 500 ml/min. Anesthetized pigs (Domestic swine, female, ∼60 kg) were attached to an activated charcoal column or a High-Flux hemodialysis filter with a blood flow rate of 200 ml/min. Animals were given an initial i.v. infusion of dabigatran (7.5 mg in 15 min) and then reduced to 5 mg/hr to achieve steady state dabigatran over 1hr. Infusion was then stopped and elimination methods were applied over 4 hrs. An observation time of 1 hr followed. Dabigatran plasma levels were quantitated with diluted thrombin time. Preliminary settings/flow rates were obtained in vitro using 5L citrated porcine whole blood exposed to different AC or HD conditions. Results: Activated charcoal completely removed dabigatran within 1 hr from the 5L whole blood supplemented with 1000 ng/mL dabigatran, with a clearance rate of 100%. By repeatedly reapplying dabigatran, it was shown the active charcoal filter had a maximum binding capacity of ∼30 mg drug. Upon saturation there was no further clearance of dabigatran. Hemodialysis removed dabigatran with increasing clearance rates depending on dialysate flow rates (100 ml/min-35%, 200 ml/min-60%, 300 ml/min-65%) reaching a plateau of ∼65%. Further increases of dialysate flow to 500 ml/min had no further effect on drug clearance. Initial plasma levels of dabigatran ranged between 200–450 ng/mL after 60 min infusion in pigs. Exposure to activated charcoal or hemodialysis (dialysate flow 300 ml/min) resulted in 75–80% reduction in circulating dabigatran after 1 hr as compared to ∼25% reduction untreated controls after 1 hr. After 2 hrs dabigatran levels were below the detection limit using both elimination methods. Conclusions: Dabigatran can effectively be removed from the circulation in this in vivo porcine model using dialysis based approaches, which results in a restoration of blood coagulation. Active charcoal perfusion was fast and effectively removed dabigatran, but may be saturated if dabigatran plasma levels are too high (human body load for 150 mg dose in steady state is ∼14g). Stationary hemodialysis with sufficiently high dialysate flows achieves similar results in this model without saturation limitations; however, the set up for dialysis is much more specialized than the simpler approach of activated charcoal filtration. Disclosures: Formella: Boehringer Ingelheim: Employment. Clemens:Boehringer Ingelheim (Anticoagulant Therapy): Employment. van Ryn:Boehringer Ingelheim: Employment. Schenk:Boehringer Ingelheim: Research Funding.

scholarly journals Direct somatic embryogenesis and plant regeneration in tea by temporary liquid immersion Embriogenesis somatik langsung dan regenerasi tanaman teh melalui perendaman sesaat

2016 ◽  
Vol 68 (1) ◽  
Author(s):  
J S TAHARDI ◽  
Tatik RAISAWATI ◽  
Imron RIYADI ◽  
W A DODD
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Culture Technique ◽  
Direct Somatic Embryogenesis ◽  
Temporary Immersion System ◽  
Single Leaf ◽  
Half Strength ◽  
Rapid Dissemination ◽  
Planting Materials

Ringkasan Perbanyakan tanaman teh [Camellia sinen­sis (L.) O. Kuntze] melalui stek tunas berdaun tunggal hanya dapat menghasilkan klon unggul dalam jumlah terbatas. Oleh sebab itu diperlukan metode alternatif dengan teknik kultur sel dan jaringan untuk perbanyakan klonal secara cepat. Dalam penelitian ini dikembangkan metode yang lebih efektif untuk regenerasi tanaman teh melalui embriogenesis somatik langsung. Massa pro­embriogenik dari eksplan kotiledon dihasilkan dengan frekuensi 56,7% dalam media MS padat setengah konsentrasi yang mengandung BAP 2 mg1L. Proliferasi, perkembangan, pendewasaan dan perkecambahan embrio somatik diperoleh dengan sistem perendaman sesaat (SPS) yang menggunakan media MS cair setengah konsen­trasi, yang diperkaya dengan zat pengatur tumbuh dengan berbagai konsentrasi. Proliferasi embrio meningkat 4,3 kali dalam media yang diberi BAP 2 mglL; perkembangan dan pendewasaannya meningkat dengan penambahan kinetin dan ABA masing-masing pada konsentrasi 0,1 mg1L yang 30% diantaranya berkecambah dan membentuk planlet tanpa penambahan zat pengatur tumbuh. Protokol SPS tersebut merupakan sistem in vitro yang berpotensi bagi proliferasi dan perkembang­an embrio somatik tanaman teh yang cepat dan sinkron dari kultur kotiledon, serta regenerasinya menjadi planlet tanpa melalui fase kalus.Summary Tea propagation by single-leaf bud cuttings has limited applications for rapid dissemination of planting materials from new elite clones. An alternative method for rapid cloning by cell and tissue culture technique is necessary. In this study we have established an improved method for tea [Camellia sinensis (L.) O. Kuntze] plant regenera­tion via direct somatic embryogenesis. Clumps of proembryogenic masses were initiated at a fre­quency of 56.7% from cotyledonary slices cul­tured on a half-strength MS agar-gelled medium supplemented with 2 mg/L BAP. Proliferation, development, maturation and germination of so­matic embryos were achieved using the temporary immersion system (TIS) provided with half­strength MS liquid media supplemented with varying concentrations of growth regulators. Em­bryo proliferation increased by 4.3-fold in me­dium provided with 2 mg/L BAP; their develop­ment and maturation were enhanced by the presence of both kinetin and ABA at 0.1 mg/L each. Germination and plant recovery were achieved at a frequency of about 30% without the use of growth regulators. The TIS protocol des­cribed above represents an in vitro system poten­tial for rapid proliferation and synchronized development of tea somatic embryos from cotyledon cultures, and their regeneration into plantlets without an intervening callus phase.

scholarly journals Micropropagation of Endemic and Endangered Mexican Species of Ponytail Palms

HortScience ◽  
2005 ◽  
Vol 40 (5) ◽  
pp. 1481-1484 ◽  
Author(s):  
María Luisa Osorio-Rosales ◽  
Martín Mata-Rosas
Keyword(s):  
Endangered Species ◽  
Growth Regulators ◽  
Activated Charcoal ◽  
Initial Treatment ◽  
Survival Rates ◽  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Ms Medium ◽  
Mexican Species

Experiments were conducted to establish an efficient protocol of micropropagation of Beaucarnea gracilis and B. recurvata two endemic and endangered Mexican species. Multiple shoots were induced by direct organogenesis from in vitro seedlings and longitudinal sections of seedlings in both species. The highest formation of shoots per explant, both B. gracilis and B. recurvata, was obtained from longitudinal sections of seedlings on Murashige and Skoog (MS) medium supplemented with 22.2 μm 6-benzylaminopurine, induced 8.2 and 11.1 shoots per explant respectively. In vitro rooting was readily achieved on MS medium with 1 g/l activated charcoal without growth regulators. According to initial treatment and depending on where the shoots come from, the rooting rates were 61% to 100% for B. gracilis, and 83% to 100% for B. recurvata. Survival rates in greenhouse conditions for both species were 80% to 100% after 3 months. These results indicate that the micropropagation of these species of Beaucarnea is technically feasible, and that in vitro culture is a useful option for the conservation and propagation of these important endangered species.

scholarly journals Somatic embryogenesis in Citrus sinensis, C. reticulata AND C. nobilis x C. deliciosa

2002 ◽  
Vol 59 (1) ◽  
pp. 41-46 ◽  
Author(s):  
Adriana Patrícia Ricci ◽  
Francisco de Assis Alves Mourão Filho ◽  
Beatriz Madalena Januzzi Mendes ◽  
Sonia Maria de Stefano Piedade
Keyword(s):  
Somatic Embryogenesis ◽  
Activated Charcoal ◽  
Somatic Embryos ◽  
Culture Medium ◽  
Sweet Orange ◽  
Mature Embryos ◽  
Indirect Somatic Embryogenesis ◽  
Kinnow Mandarin ◽  
Ponkan Mandarin

Most of the plant regeneration processes in citrus, through tissue culture, involve indirect somatic embryogenesis. The optimization of these processes is important for the development of in vitro plant improvement and micropropagation studies. Studies to evaluate the effect of different carbohydrates in somatic embryogenesis were conducted using calli from 'Ponkan' mandarin (Citrus reticulata, Blanco), 'Cravo' mandarin (C. reticulata), 'Itaboraí' sweet orange (C. sinensis L. Osbeck.), 'Valencia' sweet orange (C. sinensis) and 'Kinnow' mandarin (C. nobilis Loureiro x C. deliciosa Tenore). The culture medium used was MT supplemented with sucrose, galactose, glucose, maltose or lactose with the following concentrations of 18, 37, 75, 110, and 150 mM. The culture medium used for the maturation of somatic embryos had 0, 15, 29, 44, 58 and 73 mM of sucrose, in presence or absence of 0.5 g L<FONT FACE=Symbol>-</FONT>1 of activated charcoal. Seventy-three mM of sucrose with 0.1 mg L<FONT FACE=Symbol>-</FONT>1 of GA3 in the presence or absence 0.5 g L<FONT FACE=Symbol>-</FONT>1 of activated charcoal was also tested. Overall, the carbohydrates galactose or lactose induced a higher number of somatic embryos. Sucrose concentrations of 58 and 73 mM generated a higher number of plantlets from mature embryos of 'Ponkan' mandarin and 'Valencia' sweet orange.

scholarly journals Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

2020 ◽  
Vol 29 (2) ◽  
pp. 137
Author(s):  
Fitri Rachmawati ◽  
Dewi Permanik ◽  
Ronald Bunga Mayang ◽  
Budi Winarto
Keyword(s):  
Somatic Embryogenesis ◽  
Activated Charcoal ◽  
Embryogenic Callus ◽  
Culture System ◽  
In Vitro Propagation ◽  
Clonal Propagation ◽  
Ms Medium ◽  
Indirect Somatic Embryogenesis ◽  
Half Strength

<p>Protokol perbanyakan klonal yang efektif dan efisien sangat diperlukan untuk produksi benih berkualitas pada komersialisasi produk unggulan hasil pemuliaan. Penelitian bertujuan untuk mendapatkan protokol perbanyakan klonal Dendrobium ‘Balithi CF22-58’ melalui embriogenesis tidak langsung. Percobaan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Januari hingga Desember 2017. Penelitian ini menekankan pada penggunaan  jenis eksplan, media, dan sistem kultur. Jenis eksplan yang diuji adalah tunas pucuk, tunas lateral, dan pangkal plantlet dengan tiga media inisiasi [½ Murashige and Skoog (MS) dikombinasikan dengan 1,5 mg/l thidiazuron (TDZ) dan 0,5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2,5 mg/l metathopolin (mT) dan 0,05 mg/l BAP (MI-2), dan 5 mg/l mT dan 0,05 mg/l BAP (MI-3)]; empat media proliferasi, yaitu ½ MS dengan kombinasi: MP-1 (0,75 mg/l TDZ + 0,25 mg/l BAP), MP-2 (1,5 mg/l TDZ + 0,5 mg/l BAP), MP-3 (2,5 mg/l mT + 0,05 mg/l BAP), dan MP-4 (5,0 mg/l+ 0,05 mg/l BAP); dua sistem kultur (padat dan cair); dan tiga media regenerasi MPP-1 (½ MS dengan vitamin penuh (1/2 MS-FV) + 2% charcoal); MPP-2 (½ MS-FV); dan MPP-3 (2 g/l Rosasol 18:18:18 TE). Percobaan disusun menggunakan rancangan acak kelompok faktorial dengan lima ulangan. Hasil penelitian menunjukkan bahwa inisiasi kalus embriogenik (KE) tertinggi, yaitu 38,3% dengan waktu inisiasi 16,8 hari dihasilkan dari eksplan pangkal plantlet pada medium MI-1. Medium MP-2 dan sistem kultur cair mampu mempertahankan proliferasi KE sampai 83,1% dengan rasio penggandaan 3,23 kali. Perkecambahan embrio terbaik sampai 86,9% embrio berkecambah dengan 18,2 kecambah per rumpun dalam waktu 21,3 hari, ditunjukkan pada medium MPP-1, sedangkan pembesaran plantlet terbaik mencapai tinggi plantlet sampai 5 cm, jumlah daun hingga 4,9 helai, dan jumlah akar  2,8, dengan  2,6 cm panjang akar dan 0,27 g bobot basah plantlet, diperoleh pada medium MPP-3. Perbanyakan anggrek dengan protokol ini diperkirakan dapat menghasilkan sekitar 3.000–4.000 plantlet/eksplan/tahun. Protokol hasil penelitian ini sangat potensial diaplikasikan pada perbanyakan klonal Dendrobium melalui kultur jaringan. </p><p><strong>Keywords</strong></p><p><em>Dendrobium</em>; Embriogenesis somatik; Perbanyakan masal; Proliferasi;  Sistem kultur  </p><p><strong>Abstract</strong></p><p>The effective and efficient clonal propagation protocol is significantly needed for producing qualified seedling for commercialization of superior breeding products. The objective of the study was to establish clonal propagation protocol for Dendrobium ‘Balithi CF22-58’ via indirect somatic embryogenesis. The study was conducted at the Tissue Culture Laboratory in Indonesian Ornamental Crops Research Institute from January to December 2017. The study emphasized to utilize explant source, culture media, and culture system. Explant types were shoot tip, lateral shoot, and basal part of plantlets; three initiation media [half strength Murashige and Skoog (MS) medium containing 1.5 mg/l thidiazuron (TDZ) and 0.5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2.5 mg/l metathopolin (mT) and 0.05 mg/l BAP (MI-2), and 5 mg/l mT and 0.05 mg/l BAP (MI-3)]; four proliferation media (half strength MS medium supplemented with: MP-1 (0.75 mg/l TDZ and 0.25 mg/l BAP), MP-2 (1.5 mg/l TDZ and 0.5 mg/l BAP), MP-3 (2.5 mg/l mT and 0,05 mg/l BAP), and MP-4 (5.0 mg/l and 0.05 mg/l BAP); two culture system were solid and liquid; and three  regeneration media viz, MPP-1 (half strength MS medium with full vitamin and 2% activated charcoal); MPP-2 (MR-1 activated charcoal free), and MPP-3 (2 g/l Rosasol 18:18:18 TE). These experiments were arranged using a factorial randomized complete block design with five replications. Results of the study revealed that the highest initiation rate of embryogenic callus (EC) was up to 38.3% in 16.8 days after culture. The EC was regenerated from a basal part on MI-1 medium,  MP-2 medium and liquid culture system were able to maintain proliferation of embryogenic callus up to 83.1% with 3.23 multiplication rate. The best embryo germination up to 86.9% with 18.2 germinated embryos per clump within 21.3 days was determined on MPP-1 medium. While the best plantlet performances with 5 cm height of plantlets, 4.9 number of leaves, 2.8 number of roots, 2.6 cm root length, and 0.27 g plantlet fresh weight was obtained MPP-3 medium. With this propagation protocol, 3,000 - 4,000 plantlets/explant/year can be produced. Results of the study have high potential to be applied for in vitro propagation of Dendrobiums.</p>

scholarly journals Generation of monogenetic cattle by different techniques of embryonic cell and somatic cell cloning - their application to biotechnological, agricultural, nutritional, biomedical and transgenic research

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Maria Skrzyszowska ◽  
Marcin Samiec
Keyword(s):  
Somatic Cell ◽  
Nuclear Transfer ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Embryonic Cell ◽  
Cell Cloning ◽  
Genetic Rescue ◽  
The Family ◽  
Current Article

AbstractThe development of effective approaches for not only the in vitro maturation (IVM) of heifer/cow oocytes and their extracorporeal fertilization (IVF) but also the non-surgical collection and transfer of bovine embryos has given rise to optimizing comprehensive in vitro embryo production (IVP) technology and improving other assisted reproductive technologies (ARTs), such as cattle cloning by embryo bisection, embryonic cell nuclear transfer (ECNT) and somatic cell nuclear transfer (SCNT). The primary goal of the present paper is to demonstrate the progress and achievements in the strategies utilized for embryonic cell cloning and somatic cell cloning in cattle. Moreover, the current article is focused on recognizing and identifying the suitability and reliability of bovine cloning techniques for nutritional biotechnology, agri-food and biopharmaceutical industry, biomedical and transgenic research and for the genetic rescue of endangered or extinct breeds and species of domesticated or wild-living artiodactyl mammals (even-toed ungulates) originating from the family Bovidae.

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