Degeneration of human respiratory cell ciliary beat in monolayer cell cultures

Rautiainen, Markus; Matsune, Shouji; Yoshitsugu, Masahiko; Ohyama, Masaru

doi:10.1007/bf00179306

Cited by 17 publications

(7 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In all the collagen substrata investigated, the observed CBF decrease agreed with literature information (Rautiainen et al 1993 ;Agu et al 1999). The progressive decline in the number and frequency of ciliated cells may be explained by the loss of the cells ' threedimensional geometry upon attachment (Jorissen et al 1989).…”

Section: Cbf Studiessupporting

confidence: 90%

In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies

Agu

Jorissen

Willems

et al. 2001

Journal of Pharmacy and Pharmacology

View full text Add to dashboard Cite

The aim of this study was to establish a collagen matrix-based nasal primary culture system for drug delivery studies. Nasal epithelial cells were cultured on derivatised (Cellagen membrane CD-24), polymerised (Vitrogen gel) and fibrillar (Vitrogen film) collagen substrata. Cell morphology was assessed by microscopy. The cells were further characterised by measurement of ciliary beat frequency (CBF), transepithelial resistance (TER), permeation of sodium fluorescein, mitochondrial dehydrogenase (MDH) activity and lactate dehydrogenase (LDH) release upon cell exposure to sodium tauro-24, 25 dihydrofusidate (STDHF). Among the three collagen substrata investigated, the best epithelial differentiated phenotype (monolayer with columnar/cuboidal morphology) occurred in cells grown on Cellagen membrane CD-24 between day 4 and day 11. Cell culture reproducibility was better with Cellagen membrane CD-24 (90%) in comparison with Vitrogen gel (70%) and Vitrogen film (< 10%). TER was higher in cells grown on Vitrogen gel than on Cellagen membrane CD-24 and Vitrogen film. The apparent permeability coefficient (Papp x 10(-7)cm s(-1)) of sodium fluorescein in these conditions was 0.45+/-0.08 (Vitrogen gel) and 1.91+/-0.00 (Cellagen membrane CD-24). Except for LDH release, CBF and cell viability were comparable for all the substrata. Based on MDH activity, LDH release, CBF, TER and permeation studies, Cellagen membrane CD-24- and Vitrogen gel-based cells were concluded to be functionally suitable for in-vitro nasal drug studies. Vitrogen film-based cultures may be limited to metabolism and cilio-toxicity studies.

show abstract

Section: Cbf Studiessupporting

confidence: 90%

In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies

Agu

Jorissen

Willems

et al. 2001

Journal of Pharmacy and Pharmacology

View full text Add to dashboard Cite

show abstract

“…Because of its similarity to in vivo ciliogenesis, we used the floating ciliated culture system in this study. In a HSV system, only a thin specimen such as a monolayer cell culture on a collagen-coated cover glass can be observed clearly at high magnification [14]. However, with our method, we could observe moving cilia even on thick specimens such as a floating gel at high magnification: by attaching the convex site of the surface of the cul-tured cell layer on pieces removed from a floating cell culture to the cover glass, moving cilia could be observed on the concave site and the edge of the culture cell layer without any damage, including the movement of individual cilia (figs 1, 2).…”

Section: Discussionmentioning

confidence: 99%

Ciliary motility and surface morphology of cultured humanrespiratory epithelial cells during ciliogenesis

Yoshitsugu

Hanamure

Furuta

et al. 1994

Biology of the Cell

View full text Add to dashboard Cite

We used human respiratory epithelial cells in floating culture to study the motility and surface morphology of the cilia during ciliogenesis. Ciliary motility and surface morphology was examined by a high-speed video system and electron microscopy, respectively. On day 8 of floating culture, but rarely before, cells with numerous moving short cilia with mean ciliary beat frequency (CBF) of 17.8 +/- 2.5 Hz (mean +/- SD) were seen. The mean CBF was highest on day 10 (19.6 +/- 3.9 Hz) and reached 17.4 +/- 2.5 Hz on day 14. The ciliary beat amplitude (CBA) was recognized on day 8, abruptly increased on day 9, then continued with aging of the culture. The intracellular coordination of ciliary beats was poor on day 8 with random beat direction but improved up to day 12 when the intracellular uniform beat direction was established. However, intercellular coordination remained absent even on day 14. Results suggest that immature short cilia have a rigid beat pattern. With the elongation of the resulting whip-like movement, the ciliary beat was gradually synchronized. Although a ciliary elongation seems to be required for making intracellular coordination, other factors are involved in intercellular coordination. Also, a high CBF does not always indicate the activity of mature cilia.

show abstract

“…Epithelial cells with normal ciliary activity were not detected (87,129). Too low a cell density of the seeded nasal mucosa cells (84,142) and insufficient supplementation of the culture medium with growth-promoting factors like vitamin A (35) have been discussed as possible factors responsible for the loss of the cilia in the monolayer cultures. Moreover, the epithelial cells are detached from the basal membrane by enzymatic separation, but since this has a regulating effect on the differentiation of epithelial cells (14,68), the loss of this cell-matrix interaction can be seen as another decisive factor for the dedifferentiation of the nasal mucosa cells in the monolayer culture.…”

Section: Cell and Tissue Culture Of Human Respiratory Epitheliummentioning

confidence: 99%

Tissue Engineering in Otorhinolaryngology

Bücheler

Haisch²

2003

DNA and Cell Biology

View full text Add to dashboard Cite

Tissue engineering is a field of research with interdisciplinary cooperation between clinicians, cell biologists, and materials research scientists. Many medical specialties apply tissue engineering techniques for the development of artificial replacement tissue. Stages of development extend from basic research and preclinical studies to clinical application. Despite numerous established tissue replacement methods in otorhinolaryngology, head and neck surgery, tissue engineering techniques opens up new ways for cell and tissue repair in this medical field. Autologous cartilage still remains the gold standard in plastic reconstructive surgery of the nose and external ear. The limited amount of patient cartilage obtainable for reconstructive head and neck surgery have rendered cartilage one of the most important targets for tissue engineering in head and neck surgery. Although successful in vitro generation of bioartificial cartilage is possible today, these transplants are affected by resorption after implantation into the patient. Replacement of bone in the facial or cranial region may be necessary after tumor resections, traumas, inflammations or in cases of malformations. Tissue engineering of bone could combine the advantages of autologous bone grafts with a minimal requirement for second interventions. Three different approaches are currently available for treating bone defects with the aid of tissue engineering: (1) matrix-based therapy, (2) factor-based therapy, and (3) cell-based therapy. All three treatment strategies can be used either alone or in combination for reconstruction or regeneration of bone. The use of respiratory epithelium generated in vitro is mainly indicated in reconstructive surgery of the trachea and larynx. Bioartificial respiratory epithelium could be used for functionalizing tracheal prostheses as well as direct epithelial coverage for scar prophylaxis after laser surgery of shorter stenoses. Before clinical application animal experiments have to prove feasability and safety of the different experimental protocols. All diseases accompanied by permanently reduced salivation are possible treatment targets for tissue engineering. Radiogenic xerostomia after radiotherapy of malignant head and neck tumors is of particular importance here due to the high number of affected patients. The number of new diseases is estimated to be over 500,000 cases worldwide. Causal treatment options for radiation-induced salivary gland damage are not yet available; thus, various study groups are currently investigating whether cell therapy concepts can be developed with tissue engineering methods. Tissue engineering opens up new ways to generate vital and functional transplants. Various basic problems have still to be solved before clinically applying in vitro fabricated tissue. Only a fraction of all somatic organ-specific cell types can be grown in sufficient amounts in vitro. The inadequate in vitro oxygen and nutrition supply is another limiting factor for the fabrication of complex tissues or organ sys...

show abstract

Degeneration of human respiratory cell ciliary beat in monolayer cell cultures

Cited by 17 publications

References 8 publications

In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies

In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies

Ciliary motility and surface morphology of cultured humanrespiratory epithelial cells during ciliogenesis

Tissue Engineering in Otorhinolaryngology

Contact Info

Product

Resources

About