• Journal of Photoscience
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• 제9권2호
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• pp.21-24
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• 2002

Pineal organs of poikilotherm vertebrates transform the environmental light information into a humoral message and a neuronal activity. The former is melatonin, and the latter is modulation of the impulse in ganglion cells. The ganglion cells are physiologically classified into luminosity (achromatic) type and chromatic one, as the neural activity is modulated in two ways. We attempted to classify the pineal ganglion cells with morphological characteristics by means of the three- dimensional reconstruction method. In the pineal ganglion cells of river lamprey, there are two different features, oval and spherical. For comparison of their projection region in the brain, the tracing investigation was also carried out. The application of the neural tracer near mesencephalic tegmentum showed that only oval-shaped ganglion cells were labeled in the pineal organ. These results suggest that the oval-shaped ganglion cell is functionally different from the spherical one.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제29권4호
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• pp.249-256
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• 2003

The lack of sufficient oral mucosa available for intra-oral reconstruction has been dealt with by the use of skin or oral mucosa grafts harvested from donor sites but grafts requires more than one surgical procedures and could cause donor site morbidity. Many investigators have attempted to increase available soft tissue by tissue engineered skin or oral mucosa replacements for clinical applications. But, reconstructed mucosa by several methods have low physical properties such as rolling and contraction. The aims of this study were to develope an in vitro experimental model that maintains an epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally cultured oral mucosa embedded with Polydioxanone mesh by histological and immunohistochemical analysis. The results were as follows; 1. Oral mucosa reconstructed by three-dimensional organotypic culture revealed similar morphologic characteristics to equvalent normal oral mucosa in the point that they show stratification and differentiation. 2. The expression of cytokeratin 10/13 and involucrin in the cultured tissue showed the same pattern with normal oral mucosa suggesting that organotypic co-culture condition is able to induce cellular differentiation. 3. After insertion of polydioxanone mesh, increased tensile strength were observed. These results suggest that three-dimensional organotypic co-culture of the oral mucosa cell lines with the dermal equvalent consisting type I collagen and fibroblasts reproduce the morphologic and immunohistochemical characteristics similar to those in vivo condition. And increased physical properties by use of polydioxanone mesh will helpful for clinical applications.

• Biomolecules & Therapeutics
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• 제23권5호
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• pp.391-399
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• 2015

As a major component of the epidermal tissue, a primary keratinocyte has served as an essential tool not only for the study of pathogenesis of skin-related diseases but also for the assessment of potential toxicities of various chemicals used in cosmetics. However, its short lifespan in ex vivo setting has been a great hurdle for many practical applications. Therefore, a number of immortalization attempts have been made with success to overcome this limitation. In order to understand the immortalization process of a primary keratinocyte, several key biological phenomena governing its lifespan will be reviewed first. Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained. Finally, its application to a three-dimensional skin culture system will be described.

• 대한화장품학회지
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• 제29권2호
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• pp.105-130
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• 2003

Human skin equivalents, also designated as cultured skin substitute (Boyce and Warden, 2002) or organotypic co-cultures (Maas-Szabowski et al., 1999, 2000, 2003), are three-dimensional systems that are engineered by seeding fibroblasts into a three-dimensional dermal matrix. Such a dermal equivalent is then subsequently seeded with human keratinocytes. After cell attachment, the culture is kept first under submerged condition to allow keratinocyte proliferation. Thereafter, the culture is lifted the air-liquid interface (A/L) to expose the epidermal compartment to the air, and to further induce keratinocyte differentiation. During the air-exposure, nutrients from the medium will diffuse through the underlying dermal substrate towards the epidermal compartment and support keratinocyte proliferation and differentiation. Under these conditions, a HSE is formed that shows high similarity with the native tissue from which it was derived (Figure 1) (Bell et at., 1981; Boyce et al., 1988; Ponec et al., 1997;El Ghalbzouri et al.., 2002).

• Tissue Engineering and Regenerative Medicine
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• 제15권6호
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• pp.721-733
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• 2018

BACKGROUND: Because three-dimensional (3D) models more closely mimic native tissues, one of the goals of 3D in vitro tissue models is to aid in the development and toxicity screening of new drug therapies. In this study, a 3D skin wound healing model comprising of a collagen type I construct with fibrin-filled defects was developed. METHODS: Optical imaging was used to measure keratinocyte migration in the presence of fibroblasts over 7 days onto the fibrin-filled defects. Additionally, cell viability and growth of fibroblasts and keratinocytes was measured using the $alamarBlue^{(R)}$ assay and changes in the mechanical stiffness of the 3D construct was monitored using compressive indentation testing. RESULTS: Keratinocyte migration rate was significantly increased in the presence of fibroblasts with the cells reaching the center of the defect as early as day 3 in the co-culture constructs compared to day 7 for the control keratinocyte monoculture constructs. Additionally, constructs with the greatest rate of keratinocyte migration had reduced cell growth. When fibroblasts were cultured alone in the wound healing construct, there was a 1.3 to 3.4-fold increase in cell growth and a 1.2 to 1.4-fold increase in cell growth for keratinocyte monocultures. However, co-culture constructs exhibited no significant growth over 7 days. Finally, mechanical testing showed that fibroblasts and keratinocytes had varying effects on matrix stiffness with fibroblasts degrading the constructs while keratinocytes increased the construct's stiffness. CONCLUSION: This 3D in vitro wound healing model is a step towards developing a mimetic construct that recapitulates the complex microenvironment of healing wounds and could aid in the early studies of novel therapeutics that promote migration and proliferation of epithelial cells.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권4호
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• pp.264-268
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• 2001

Aggregates of specific cells are often regarded as better from in artificial organs and mammalian cell bioreactors in terms of cell-specific functionality. In this study, the morphology and liver specific functions of freshly harvested primary rat hepatocytes, which were cultivated as spheroids and entrapped in a synthetic thermo-reversible extracellular matrix, were examined and compared to a control (hepatocytes in single cell form). A copolymer of N-isopropylacrylamide(98 mole % in feed) and acrylic acid (poly (NiPAAm-co-AAC)), a thermo- reversible copolymer gel ma- trix, was used to entrap hepatocytes either in spheroids or single cells. During a 7-day culture pe-riod, the spheroids maintained higher viability and produced albumin and urea at a relatively con-stant rate, while, the single cell culture showed a slight increase in cell numbers and a reduction in albumin secretion Hepatocytes cultrured as spheroids present a potentially useful three-dimensional cell culture system for application in bioartificial liver device.

• 미생물학회지
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• 제22권1호
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• pp.19-28
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• 1984

A. conoides에 의한 선충 포획과정을 SEM과 TEM을 이용하여 관찰하였다. 1. A. conoides는 점착성 three-dimensional networks에 의해 선충을 포획한다. 2. Trap cell은 영양균사에 비해 세포벽이 두꺼우며 endoplasmic reticulum, mitochondria 및 electron-dense granule이 풍부하다. 3. 포획기관에서만 관찰되는 전자밀도가 높은 과립은 포획기관이 선충을 점착하여 침투하는 과정에서 소실된다. 4. 포획기관과 선충이 부착된 사이에서 osmiophilic layer가 관찰되었고 바로 이 지점으로부터 침투가 일어나며, 한 포획기관에서 두 군데 이상 동시 침투가 가능하다. 5. 포획기관의 선충내 침투시 appressorium이 형성되지 않고 침투되는 경우가 있다. 6. 균주와 선충의 혼합배지를 2~3주 두었을 때, 유충들이 포자에 접착되어 죽는다.

• International Journal of Stem Cells
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• 제17권2호
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• pp.158-181
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• 2024

This study offers a comprehensive overview of brain organoids for researchers. It combines expert opinions with technical summaries on organoid definitions, characteristics, culture methods, and quality control. This approach aims to enhance the utilization of brain organoids in research. Brain organoids, as three-dimensional human cell models mimicking the nervous system, hold immense promise for studying the human brain. They offer advantages over traditional methods, replicating anatomical structures, physiological features, and complex neuronal networks. Additionally, brain organoids can model nervous system development and interactions between cell types and the microenvironment. By providing a foundation for utilizing the most human-relevant tissue models, this work empowers researchers to overcome limitations of two-dimensional cultures and conduct advanced disease modeling research.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.141-144
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• 2000

Highly open porous polymer matrices are required for high density cell seeding, efficient nutrient, and oxygen supply to the cells cultured in the three dimensional matrices. However, there are severe problems of mass transfer limitations within the cell/scaffolds culture system. Thus we hypothesize that continuos-flow culture conditioning of cells with the scaffolds may improve the cell viability and the differentiated function. In this study, we fabricated porous PLGA scaffolds by using gas-foaming/salt-leaching method as previous described. Viscous PLGA gel paste contains ammonium bicarbonate particulates, acting as a gas-foaming agent as well as a salt-leaching porogen, were cast into Teflon mold and dried. Ammonium bicarbonate salt upon contact to an acidic aqueous solution evloves gaseous ammonia and carbon dioxide by itself. And we conjugated galactose moiety [AGA; $N-(aminobuty1)-O-{\beta}-D-galactopyranosyl-(1{\rightarrow}4)-D-glucoamide]$ to the terminal end group of a PLGA to increase the cell adhesion and matain the differentiated function of hepatocytes. Cell-seeded scaffolds were secured in a flow bioreactor chamber and exposed to continuous flow at 5 ml/min. As a result of our study, the high yield of hepatocytes attachment was accomplished by increasing the concentration of PLGA-AGA conjugate in polymer scaffolds and cells in the scaffolds under continuos flow condition maintained a high level of viability and albumin secretion rate of cultured hepatocytes showed a higher level that of control groups.

• Molecules and Cells
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• 제40권2호
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• pp.117-122
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• 2017

Despite the fact that porcine embryonic stem cells (ESCs) are a practical study tool, in vitro long-term maintenance of these cells is difficult in a two-dimensional (2D) microenvironment using cellular niche or extracellular matrix proteins. However, a three-dimensional (3D) microenvironment, similar to that enclosing the inner cell mass of the blastocyst, may improve in vitro maintenance of self-renewal. Accordingly, as a first step toward constructing a 3D microenvironment optimized to maintain porcine ESC self-renewal, we investigated different culture dimensions for porcine ICM-derived cells to enhance the maintenance of self-renewal. Porcine ICM-derived cells were cultured in agarose-based 3D hydrogel with self-renewal-friendly mechanics and in 2D culture plates with or without feeder cells. Subsequently, the effects of the 3D microenvironment on maintenance of self-renewal were identified by analyzing colony formation and morphology, alkaline phosphatase (AP) activity, and transcriptional and translational regulation of self-renewal-related genes. The 3D microenvironment using a 1.5% (w/v) agarose-based 3D hydrogel resulted in significantly more colonies with stereoscopic morphology, significantly improved AP activity, and increased protein expression of self-renewal-related genes compared to those in the 2D microenvironment. These results demonstrate that self-renewal of porcine ICM-derived cells can be maintained more effectively in a 3D microenvironment than in a 2D microenvironment. These results will help develop novel culture systems for ICM-derived cells derived from diverse species, which will contribute to stimulating basic and applicable studies related to ESCs.