scholarly journals Verification of the Field Productivity of Rehmannia glutinosa (Gaertn.) DC. Developed Through Optimized In Vitro Culture Method

Plants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 317
Author(s):  
Yong-Goo Kim ◽  
Richard Komakech ◽  
Dae Hui Jeong ◽  
Yun mi Park ◽  
Tae Kyoung Lee ◽  
...  
Keyword(s):  
Near Infrared ◽  
Chemical Constituents ◽  
Culture Method ◽  
Poor Quality ◽  
Culture Conditions ◽  
Rehmannia Glutinosa ◽  
Sterile Culture ◽  
Perennial Plant ◽  
The Family

Rehmannia glutinosa (Gaertn.) DC is a perennial plant belonging to the family Scropulariidae. The root of R. glutinosa is used in oriental medicine and mainly grown using rootstock rather than seed cultivation, which gives rise to several problems including root rot, and results in a low productivity and poor quality. To solve the challenges involved in R. glutinosa seed cultivation, our team previously used the formative features and genetic analysis of R. glutinosa to determine the optimal in vitro tissue culture conditions for producing sterile culture seedlings and rootstocks of R. glutinosa. The aim of the present study was to identify differences between R. glutinosa standard rootstock seedlings (SR), R. glutinosa culture rootstock seedlings (CR), and culture seedlings (CS) under field conditions. The reproductive characteristics of the aerial part were more robust while the area and length of leaves were smaller for SR than those for CR and CS. The characteristic that differed the most in SR was flowering, which did not occur in CR and CS. In addition, the fresh and dry weights of the subterranean parts of CR and CS were two-fold greater than those of SR. Fourier transform near-infrared (FT-NIR) analysis showed only slight differences between the chemical constituents of SR and its culture products, which was confirmed by measuring the content of catalpol, an indexing substance. Catalpol had a reduced content in the culture products compared to SR. However, this difference was not significant. Our findings will be useful for the identification of the best seedling type of R. glutinosa to enable its mass production.

scholarly journals Verification of the Field Productivity and Bioequivalence of a Medicinal Plant (Polygonum multiflorum) Developed Using an In Vitro Culture Method

Plants ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1280
Author(s):  
Yong-Goo Kim ◽  
Richard Komakech ◽  
Dae Hui Jeong ◽  
Kwonseok Jeon ◽  
Yunmi Park ◽  
...  
Keyword(s):  
Near Infrared ◽  
Culture Method ◽  
Culture Technique ◽  
Oriental Medicine ◽  
Polygonum Multiflorum ◽  
Perennial Plant ◽  
Plant Parts ◽  
Beneficial Effects ◽  
Root Tissues

Polygonum multiflorum Thunb. is a perennial plant that belongs to Polygonaceae. Root tissues are the main plant parts used as medicinal herbs in Korean oriental medicine. The P. multiflorum tuber is well known for its medicinal properties in Korean oriental medicine, and it contains a number of useful substances (secondary metabolites of emodin, 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (TSG), etc.) that are increasing in demand, as several studies show that they have beneficial effects on the human body. In this study, the production volumes and useful material content differences between cultured P. multiflorum seedlings (culture seedlings: CSs), which had been grown using a tissue culture technique under optimized conditions, and existing varieties in circulation (seed seedlings: SSs) were determined using a long-term field test. The growth characteristics of the underground parts were investigated by harvesting the tuberous roots (medicinal parts) after 1 year, and the results showed that the fresh and dry weights of the CS tubers were higher than those of the SS tubers. However, the SS rootlets had higher fresh and dry weights than the CS rootlets. A liquid chromatography-mass spectrometry component analysis of the P. multiflorum tubers and a Fourier transform near-infrared spectrophotometer analysis of the roots were undertaken. The results showed that the levels of TSG, which is a medicinal substance produced by P. multiflorum, were higher in the CSs than in the SSs, but the differences were not significant. The CS results from this study will inform future studies on the mass production of P. multiflorum in the field because the medicinal area was greater in CSs than in SSs.

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 Enhancement of chemical constituents in hydrosol and residual water of Aquilaria malaccensis tetraploid

2018 ◽  
Author(s):  
Siti Suhaila A Rahman ◽  
Keyword(s):  
Chemical Constituents ◽  
Water Soluble ◽  
Residual Water ◽  
Planting Material ◽  
Aquilaria Malaccensis ◽  
The Family ◽  
Material Choice ◽  
By Products ◽  
Better Than

Aquilaria malaccensis is an agarwood-producing species in the family Thymeleaeceae. Agarwood is a fragrant resin used in the manufacture of incense sticks, and in pharmaceutical, perfumery and cosmetic industries. In addition to the resin, hydrosol and residual water by-products from agarwood woodchip distillation are also utilized. Hydrosol contains water-soluble fragrant chemicals used as a tonic drink, in cooking and cosmetics while the residual water is used in spas and aromatic bath treatments. The present study was conducted to identify and compare compounds present in hydrosol and residual water by-products of diploid and polyploid A. malaccensis. Four different four-month-old A. malaccensis plants were compared: soil-grown diploid seedlings (DS), in vitro-grown seedlings (DV), tissue culture-derived plantlets (DC) and artificially induced tetraploid plantlets (TC). Hydrosol water from TC leaf and root samples were found to contain higher amounts of compounds compared with other samples. The TC leaf samples were qualitatively better as key compounds of agarwood such as α- and γ-eudesmol were detected. TC stem samples also contained higher amounts of key compounds compared with other samples, while the overall amount of compounds was highest in DS stem samples. The residual water of TC stem and root samples contained key compounds not detected in other samples, while DS residual water samples contained the highest total amount of compounds. Aquilaria malaccensis tetraploids performed better than their diploid counterparts in production of compounds, and thus may be a better planting material choice for commercial plantations.

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.

scholarly journals Optimization of culture conditions for in vitro adventitious roots and selected flavonoids production in Boesenbergia rotunda liquid suspension culture

Food Research ◽  
2020 ◽  
Vol 4 (S5) ◽  
pp. 26-33
Author(s):  
K.A. Ghani ◽  
A. Yusuf ◽  
N. Khalid
Keyword(s):  
Adventitious Roots ◽  
Ph Value ◽  
Culture Conditions ◽  
Inoculum Density ◽  
Root Production ◽  
Initial Inoculum ◽  
Perennial Plant ◽  
Liquid Suspension ◽  
Boesenbergia Rotunda

Boesenbergia rotunda (L.) Mansf. is one of the unique monocotyledonous perennial plant species belonging to the ginger (Zingiberaceae) family. Locally known as ‘Temu Kunci’ in Malaysia and Indonesia, this medicinal plant has been widely used in Asian dishes, particularly as a condiment or as traditional natural medicines. The important medicinal properties of B. rotunda majorly derived from flavonoids which are highly sought as pharmaceuticals. In this study, culture conditions for the growth of adventitious roots in liquid suspension cultures were optimized. The highest adventitious root production was achieved when cultured with initial inoculum density of 1.5 g and pH value at 5.8 after five weeks of culture. HPLC analysis discovered that production of valuable flavonoid compounds (pinostrobin, cardamonin and panduratin A) was significantly higher when the adventitious roots were cultured with initial inoculum density of 1.5 g whereas the initial pH medium did not significantly affect flavonoid production.

scholarly journals THE CHEMICAL CONSTITUENTS' OF THE LEAVE ESSENTIAL OIL OF ALTERNANTHERA PUNGENS (KUNTH)

2020 ◽  
Vol 10 ◽  
pp. 123
Author(s):  
Abdulrazaq Omotunde Ogunmoye ◽  
Odunayo Christy Atewolara-Odule ◽  
Oseyemi Omowunmi Olubomehin ◽  
Segun Ajibola Ogundare ◽  
Sodiq Tolulope Yussuf
Keyword(s):  
Essential Oil ◽  
Chemical Constituents ◽  
Gas Chromatography Mass Spectrometry ◽  
Traditional Use ◽  
Perennial Plant ◽  
The Family ◽  
Hexahydrofarnesyl Acetone ◽  
Leaf Oil ◽  
Compositional Pattern ◽  
The Common

Alternanthera pungens Kunth commonly called khaki weed is from the family Amaranthaceae. It is a herbaceous perennial plant that has stems prostrate, rarely rising and about 10-50 cm long. The work was carried out due to the scarcity of information on the volatile constituents from the plant leaves despite works on the flower and other parts. The extraction of the essential oils from the dried leaves was carried out by the hydro distillation method using an all-glass Clevenger apparatus. The extracted oils were then analyzed using gas chromatography-mass spectrometry (GC-MS). A total of twelve constituents' representing 93.39% of A. Pungens oil with a yield of 0.4% (v/w) was obtained. The analysis of the GC-MS results of the leaf oil showed that it was dominated by â-ionone (42.18%) and hexahydrofarnesyl acetone (15.53%), others in trace amounts include; methyl palmitate (6.13%), 1octadecyne (4.72%), undecane (3.73%), para-mentha-1, 3, 8-triene (3.65%), isophytol (3.21%), ?cadinene (3.06%), 1, 2-dimethyl cyclooctene (3.05%), para-cymene (2.96%), phytol (2.67%) and neophytadiene (2.50%). The  common classes of compounds present in the leaves oil are aceto monocyclic monoterpenoid (42.18%), sesquiterpenoids (18.59%), hydrocarbons (11.50%), diterpenoids (8.38%), monoterpenes (6.61%) and fatty acids (6.13%).The constituents and the compositional pattern of essential oil identified from the leaves of Alternanthera pungens grown in Nigeria differ quantitatively and qualitatively from previously reported member of the genus and the presence of sesquiterpenoid as one of the major components of the oils justify the traditional use of the plants in treating pains, headaches and inflammations.

scholarly journals An efficient in vitro propagation protocol of Dianthus giganteiformis Borbas subsp. kladovanus (Degen) Soo

2018 ◽  
pp. 77-85
Author(s):  
Marija Markovic ◽  
Mihailo Grbic ◽  
Matilda Djukic
Keyword(s):  
In Vitro Propagation ◽  
Sand Dunes ◽  
Germination Percentage ◽  
Plant Production ◽  
Ms Medium ◽  
Sterile Culture ◽  
Perennial Plant ◽  
Danube Region

Dianthus giganteiformis subsp. kladovanus is an endemic, endangered, horticulturally appealing perennial plant that can be used for the revegetation of sand dunes of the Danube region. The appropriate method for its effective production is micropropagation. For this reason, the experiments were conducted in order to establish an efficient protocol for the micropropagation of this subspecies. The sterile culture was initiated from seeds collected in situ and the germination percentage was high (88%). According to the results obtained in this study, the multiplication phase should be performed on an MS medium enriched with 0.1 mg/L BAP and 0.1 mg/L NAA. The concentration of MS salts significantly influenced rooting, and higher rooting percentages were obtained on reduced MS media (91.7 - 95%) than on MS media (73.4 - 76.7%). The addition of NAA slightly increased rooting percentage (up to 95%). Obtained microplants were successfully acclimatized (83.3%) in a substrate composed of peat and sand (1: 1; v/v). Using the protocol presented in this paper, the efficient propagation of D. giganteiformis spp. kladovanus can be achieved for rapid plant production aimed at revegetation, biodiversity protection and floricultural production of this species.

scholarly journals Development of an in vitro tissue culture system for hammer coral (Fimbriaphyllia ancora) ovaries

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yi-Ling Chiu ◽  
Ching-Fong Chang ◽  
Shinya Shikina
Keyword(s):  
Structural Integrity ◽  
Culture Method ◽  
Culture Conditions ◽  
Short Term ◽  
Wide Range ◽  
Fetal Bovine ◽  
Short Term Culture ◽  
Mesenterial Filaments

AbstractIn vitro gonad culture systems have proven useful to investigate intrinsic mechanisms of sexual reproduction in animals. Here we describe development of an in vitro culture method for coral ovaries. Mesenterial tissues containing both ovaries and mesenterial filaments were microscopically isolated from the scleractinian coral, Fimbriaphyllia ancora, and culture conditions were optimized. M199 diluted 10× (10% M199, pH 8.1) and supplemented with 25 mM HEPES and the antibiotics, ampicillin, penicillin and streptomycin, supported oocyte survival and maintained the structural integrity of ovaries during short-term culture (~ 6 days). Addition of a commercial antibiotic–antimycotic solution (Anti–Anti) and fetal bovine serum adversely affected ovary maintenance and caused tissue disintegration. Characterization of cultured ovaries showed that there is no difference in cell proliferation of ovarian somatic cells between culture Days 1 and 6. Moreover, the presence of oogonia and expression of a major yolk protein, vitellogenin, were confirmed in ovaries cultured for 6 days. This system will be useful for studying effects of a wide range of substances on coral oogenesis.

P–216 Successful pregnancies and deliveries in patients with a recurrent failure of ART treatments following artificial removal of the zona pellucida (ZP) at the pronuclear stage

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Mio ◽  
K Yumoto ◽  
T Shimura ◽  
M Sugishima ◽  
M Nakaoka ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Culture Method ◽  
Oocyte Retrieval ◽  
Time Lapse ◽  
Poor Quality ◽  
Blastocyst Transfer ◽  
Human Embryos ◽  
Culture Methods ◽  
Conventional Culture

Abstract Study question Can a novel embryo culture method that artificially removes the ZP at the pronuclear stage yield successful pregnancy in patients with poor-quality embryos and/or blastocysts? Summary answer A blastocyst transfer after ZP-free culture can result in pregnancy for patients who cannot obtain good quality blastocysts from conventional culture methods. What is known already Perivitelline threads are been associated with the formation of cytoplasmic fragments. We had previously observed perivitelline threads in the adhesive region between the ooplasm and the ZP at the first cleavage in human embryos. We removed the ZP at the pronuclear stage in 71 abnormally fertilized oocytes (zygotes with three pronuclei), donated after conventional IVF (c-IVF), and termed them ZP-free 3PN. We found ZP-free 3PN embryos could be cultured without losing blastomere adhesions. Furthermore, the rate of good quality embryos was significantly higher in ZP-free 3PN embryos compared with ZP-intact embryos (ZP-intact 2PN/2PB and 3PN embryos; P < 0.05). Study design, size, duration This study was conducted in two cases selected among patients who underwent ART treatment in our clinic between 2018 and 2019. Cases were selected if they lacked good quality blastocysts in previous c-IVF/Intracytoplasmic Sperm Injection (ICSI) cycles due to massive cytoplasmic fragmentation at the first and second cleavage. We performed a clinical trial of ZP-free culture from December 2019 to March 2020. Participants/materials, setting, methods Two cases were selected for this trial. Normally fertilized oocytes were grouped as ZP-free or ZP-intact. For the ZP-free group, 2PN embryos were placed in 0.125M sucrose-containing HEPES to reduce ooplasm size, then ooplasms were completely separated from ZPs by a laser and pipetting. ZP-free and ZP-intact embryos were cultured with time-lapse imaging for up to seven days. Resultant blastocysts were either transferred into uterus or cryopreserved on Day5/6/7 for future embryo transfer cycles. Main results and the role of chance The ZP-free culture method was applied to two patients (patient A and B) with recurrent failure of ART in our clinic due to poor-quality embryos and/or difficulties in obtaining good quality blastocysts. In both cases, blastocysts were successfully obtained and cryopreserved for all ZP-free culture cycles. In patient A, one good quality ZP-free blastocyst was freshly transferred five days after oocyte retrieval, and a live male baby (2925g) was delivered at 40 weeks of gestation by caesarean section). In patient B, a frozen/thawed ZP-free blastocyst transfer was conducted, and a live female baby (3225g) was delivered at 39 weeks of gestation by vaginal delivery. This shows ZP-free culturing may help obtain viable embryos in patients for which conventional in vitro culturing methods result in embryos characterized with severe cytoplasmic fragmentation and poor quality in the early cleavage stage. Limitations, reasons for caution Although successful pregnancies and deliveries were confirmed in two cases, postnatal evaluations will be absolutely necessary for infants derived from ZP-free culture. In addition, the number of trial cases needs to be expanded, however careful selection of suitable patients is necessary for this novel culture method. Wider implications of the findings: We found removing the ZP at the pronuclear stage improved embryo development and led to successful pregnancies and deliveries after blastocyst transfer. This indicates ZP-free culturing may be an effective method for decreasing cytoplasmic fragmentation caused by perivitelline threads or adhesion between the ooplasm and the zona pellucida. Trial registration number Not applicable

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.

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