• Title/Summary/Keyword: tissue-culture

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Several Factors on Bulblets Regeneration from Callus Culture in Lilium longiflorum 'Celia' (백합 'Gelia' 캘러스로부터 자구 재분화에 미치는 제요인)

  • 박소영;김시동;신세균;이철희;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.183-188
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    • 1997
  • Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

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Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model

  • Thuwanut, Paweena;Comizzoli, Pierre;Pimpin, Alongkorn;Srituravanich, Weerayut;Sereepapong, Wisan;Pruksananonda, Kamthorn;Taweepolcharoen, Charoen;Tuntiviriyapun, Punkavee;Suebthawinkul, Chanakarn;Sirayapiwat, Porntip
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.2
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    • pp.111-123
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    • 2021
  • Objective: Using domestic cats as a biomedical research model for fertility preservation, the present study aimed to characterize the influences of ovarian tissue encapsulation in biodegradable hydrogel matrix (fibrinogen/thrombin) on resilience to cryopreservation, and static versus non-static culture systems following ovarian tissue encapsulation and cryopreservation on follicle quality. Methods: In experiment I, ovarian tissues (n=21 animals; 567 ovarian fragments) were assigned to controls or hydrogel encapsulation with 5 or 10 mg/mL fibrinogen (5 or 10 FG). Following cryopreservation (slow freezing or vitrification), follicle viability, morphology, density, and key protein phosphorylation were assessed. In experiment II (based on the findings from experiment I), ovarian tissues (n=10 animals; 270 ovarian fragments) were encapsulated with 10 FG, cryopreserved, and in vitro cultured under static or non-static systems for 7 days followed by similar follicle quality assessments. Results: In experiment I, the combination of 10 FG encapsulation/slow freezing led to greater post-thawed follicle quality than in the control group, as shown by follicle viability (66.9%±2.2% vs. 61.5%±3.1%), normal follicle morphology (62.2% ±2.1% vs. 55.2%±3.5%), and the relative band intensity of vascular endothelial growth factor protein phosphorylation (0.58±0.06 vs. 0.42±0.09). Experiment II demonstrated that hydrogel encapsulation promoted follicle survival and maintenance of follicle development regardless of the culture system when compared to fresh controls. Conclusion: These results provide a better understanding of the role of hydrogel encapsulation and culture systems in ovarian tissue cryopreservation and follicle quality outcomes using an animal model, paving the way for optimized approaches to human fertility preservation.

The Relationship between Body Mass Index(BMI), Adipocyte Size and Leptin and Angiotensin II Secretion in Human Adipose Tissue

  • Heo Young-Ran;Moustaid-Moussa Naima
    • Journal of Community Nutrition
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    • v.8 no.2
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    • pp.69-75
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    • 2006
  • Adipose tissue has now been recognized as a rich source of metabolically active molecules that include leptin and angiotensinogen (AGT), the precursor of angiotensin II (Ang II). Both of which have been implicated in the pathogenesis of metabolic alteration and hypertension associated with obesity. In this study, we examined the relationship between body mass index (BMI), adipocyte size, leptin, Ang II secretion and mRNA expression in human adipose tissue obtained from female subjects. Leptin and Ang II were analyzed using specific radioimmunoassay kits following a 48hour tissue culture. Leptin and Ang II secretion varied from 1.4 - 72.1ng/g and 0.8 - 57.3pg/g of tissue respectively. These large individual variations limit significant correlation between BMI, leptin and Ang II secretion. Ang II secretion was significantly higher in the obese than the non-obese (p < 0.05) and positively correlated with BMI. However, no difference in leptin secretion between the obese and the non-obese was observed and leptin secretion showed negative correlation with BMI. No difference in leptin and AGT mRNA expression in adipose tissue between the obese and the non-obese was observed. Although several limitations of this study, we found increased Ang II secretion in obese patients compared with non-obese patients, and positive correlation between AGT and BMI. Observed difference in AGT expression between the obese and the non-obese in this study might be of importance in relation with obesity related hypertension. (J Community Nutrition 8(2): 69-75, 2006)

Cross-Linked Collagen Scaffold from Fish Skin as an Ideal Biopolymer for Tissue Engineering

  • Biazar, Esmaeil;Kamalvand, Mahshad;Keshel, Saeed Heidari;Pourjabbar, Bahareh;Rezaei-Tavirani, Mustafa
    • Korean Journal of Materials Research
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    • v.32 no.4
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    • pp.186-192
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    • 2022
  • Collagen is one of the most widely used biological materials in medical design. Collagen extracted from marine organisms can be a good biomaterial for tissue engineering applications due to its suitable properties. In this study, collagen is extracted from fish skin of Ctenopharyngodon Idella; then, the freeze drying method is used to design a porous scaffold. The scaffolds are modified with the chemical crosslinker N-(3-Dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) to improve some of the overall properties. The extracted collagen samples are evaluated by various analyzes including cytotoxicity test, SDS-PAGE, FTIR, DSC, SEM, biodegradability and cell culture. The results of the SDS-PAGE study demonstrate well the protein patterns of the extracted collagen. The results show that cross-linking of collagen scaffold increases denaturation temperature and degradation time. The results of cytotoxicity show that the modified scaffolds have no toxicity. The cell adhesion study also shows that epithelial cells adhere well to the scaffold. Therefore, this method of chemical modification of collagen scaffold can improve the physical and biological properties. Overall, the modified collagen scaffold can be a promising candidate for tissue engineering applications.

Studies on tissue culture of medicinal plants (II) -Tissue cultures of Glycyrrhiza glabra L. var. glandulifera $R_{EG.}$ et $H_{ERDER}$- (약용식물(藥用植物)의 조직배양(組織培養)에 관(關)한 연구(硏究)(II) -이태리감초(甘草)의 조직배양(組織培養)-)

  • Yoo, Sung-Cho;Kim, Sung-Soon
    • Korean Journal of Pharmacognosy
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    • v.7 no.1
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    • pp.55-57
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    • 1976
  • The callus formation of Glycyrrhiza glabra L. var. $glandulifera\;R_{EG}.\;et\;H_{ERD}$. in tissue culture was promoted on Murashige and Skoog's basal solution supplemented with 40g/l of sucrose, 1mg/l of kinetin and 5mg/l of 2, 4-D. The fresh and dry weights of callus and glycyrrhizin contents in callus of the Glycyrrhiza glabra L. var. $glandulifera\;R_{EG}.\;et\;H_{ERD}$. were determined monthly up to 12 months and obtained the results as follows: 1.The fresh weight of formed callus was increased rapidly from 2 to 4 months but growing rate of callus was slow from 4 to 6 months. This indicates that the cell division of callus was most active during the first $2{\sim}3$ months. 2. Glycyrrhizin contents in callus were also increased but the contents were not related to the increased weight of callus.

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Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells (미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구)

  • Song, Jung-Ho;Park, Bong-Wook;Byun, June-Ho;Kang, Eun-Ju;Rho, Gyu-Jin;Shin, Sang-Hun;Kim, Uk-Kyu;Kim, Jong-Ryoul
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.3
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    • pp.186-196
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    • 2010
  • Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

Production Method of Basic Seed Potatoes (Solanum tuberosum L.) by Stem Cuttings, Microtuberization and Hydroponic Culture (莖揷, 器內小塊莖形成, 養液裁培에 의한 감자 無病株 생산 方法)

  • 김현준;김숭열;신관용;김학기;김화영
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.83-86
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    • 1997
  • To produce and supply the healthy potato, basic seed potatoes were produced by stem cutting, microtuberization and hydroponic culture. The total number of tubers and the total tuber weight per $\textrm{m}^2$ of potato were more in hydroponic culture as each products were 1, 152 and 4, 492g than in the stem cutting (75 and 4, 136g) or microtuberization (1, 080 and 1, 080g) using petridishes. The total yield per 10a in the field was propagated highly stem cutting > propagated microtubers > hydroponics > microtubers. The number of tubers per 10a produced by hydroponics (33, 064) was higher than any other methods. This indicated the hydroponic culture can be used in the multiplication of basic seed potatoes.

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Multiple Shoot Induction from Ex Vitro and In Vitro Derived Stein Node Culture of Populus alba L.$\times$P.grandidentata Michx. (줄기 절간조직 배양에 의한 교잡종 사시나무의 대량증식)

  • Sung Ho SON;Richard B. HALL
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.131-135
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    • 1995
  • Physiologically modified stem nodes derived from ex vitro and in vivo explants of hybrid aspen (Populus alba L.X P.grandidentata Michx. 'Crandon') were tested for their multiple shoot regeneration capacity using a broad spectrum dosage of cytokinins. Ex vitro derived stem nodes with excised axillary buds at the time of culture produced 11 to 13 multiple shoots on 20 to 30 $\mu$M zeatin containing Woody Plant Medium (WPM) after 6 weeks. Excision of axillary bud sprouts after 2 weeks of culture and culture of the remaining stem nodes on WPM with 1.0 to 2.0 $\mu$M BA or 10 to 30 $\mu$M zeatin produced 13 to 15 and 7 to 8 shoots per explant, respectively, Multiple tiny shoots were produced when in vivo derived stem nodes (on which all leaves were removed) were cultured on WPM with 30 to 50 $\mu$M 2iP or 20 to 50 $\mu$M zeatin. The greatest number of multiple tiny shoot proliferation (32 to 50 shoots per explant) were obtained when the explants were cultured on media containing 20 $\mu$M zeatin. Successful transplanting of these multiple shoots into the greenhouse and/or nursery was achieved.

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Varietal Difference in Plant Regeneration from Cotyledon Culture of Capsicum annuum L. (고추 자엽에서 식물체 재분화의 품종간 차이)

  • 오명규;이영만;박문수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.301-304
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    • 1998
  • Effects of genotype and culture medium on plant regeneration from cotyledon segments of red pepper(Capsicum annuum L.) was investigated. Among combinations of IAA(0.25 and 0.50 mg/L) and zeatin(2.0 and 4.0 mg/L) added to MS medium, combination of 2.0 mg/L zeatin and 0.25 mg/L IAA was shown to be the best for shoot differentiation from cotyledon segments. Shoot regeneration from cotyledon explants took 9 to 25days, depending on genotypes and culture media. Early shooting was observed in Yeongyangjaelae, Putgochw, Karkovskij-A-35, Gris I-A-1 on MS medium containing 2.0 mg/L zeatin and 0.25 IAA mg/L. Percent of explants producing shoots, as also influenced by genotypes and culture media, were over 90% for 621, Yeongyangjaelae, Putgochw, Nikko jacksacgmulgochw, Ch-6-Num-216, and Kajenskij-A-35 when cultured on MS medum supplemented with 2.0 mg/L zeatin and 0.25 mg/L IAA and for Fresno chile, PI 169126, Kajenskij-A-35, jacksacgmulgochw, and PI 297438 on MS medium including 2.0 mg/L BA and 1.0 mg/L IAA.

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Mass Propagation of Vitex negundo L., in vitro

  • Thiruvengadam, Muthu;Jayabalan, Narayanasamypillai
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.151-155
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    • 2000
  • Shoot proliferation was obtained from shoot tips and nodal explants of Vitex negundo L. on MS medium supplemented with either BAP or KIN (0.1-2.0 mg/L) alone or in combination with NAA (0.1 mg/L). The concentrations of cytokinins combined with NAA produced multiple shoots from shoot tips and nodal explants. The highest mean percentage (84.3$\pm$8.0) of shoot multiplication's were observed on nodal explants in the presence of BAP (1.5 mg/L) and NAA (0.1 mg/L) followed by shoot tips (65.0$\pm$5.0). The regenerated shootlets were rooted on MS basal medium IAA, IBA, NAA (0.1-1.5 mg/L). The maximum number of roots (51.0$\pm$2.6) was achieved on the medium containing IBA (1.0 mg/L) followed by other auxins (NAA, IAA). The regenerated plants were successfully transferred to a mixture of vermiculate and soil. About 95% of the plantlets survived when transferred to the field.

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