Growth Pattern of Sirc Rabbit Corneal Cells in Microwell Inserts

Hutak, Christine M.; Kavanagh, Marie E.; Reddy, Indra K.; Barletta, Michael

doi:10.3109/15569529709048892

Cited by 16 publications

(17 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, no biologically significant differences in the morphology of the cells were noted on any day sampled. The number of cell layers formed on day 10 in culture was similar to the growth pattern we had seen in the original growth study [6].…”

Section: Discussionsupporting

confidence: 65%

“…We have previously demonstrated the multiple cell layer growth pattern of SIRC rabbit corneal cells on polycarbonate-based filter inserts [6] and have used SIRC rabbit corneal cells in a model to evaluate corneal epithelial permeability [5]. These studies revealed a growth pattern similar to that of rabbit corneal epithelium [6] and an electrical resistance similar to that of the rabbit cornea [5].…”

Section: Introductionmentioning

confidence: 98%

See 1 more Smart Citation

Comparative Development of SIRC Rabbit Corneal Cells Grown on Polycarbonate- and Polyester-Based Filters

Hutak

Kavanagh

Reddy

2002

Skin Pharmacol Physiol

Self Cite

View full text Add to dashboard Cite

Statens Seruminstitut Rabbit Cornea (SIRC) cells were grown on sterile polycarbonate- and polyester-based filter inserts to evaluate the feasibility of substituting the polyester-based filter inserts for use in a previously developed permeability model. The two filter types were compared for differences in cellular growth rate and morphological changes following days 3, 7 and 10 in culture. There were no significant differences in the number of cell layers formed on any day examined. The numbers of cell layers formed with the polycarbonate-based filter inserts were 1.75 ± 0.09, 2.53 ± 0.14 and 2.93 ± 0.16, for days 3, 7 and 10, respectively. The numbers of cell layers formed with the polyester-based filter inserts were 1.79 ± 0.07, 2.23 ± 0.11 and 2.90 ± 0.13, for days 3, 7 and 10, respectively. The SIRC cells had a similar comparative morphology on each of the above days. Active cell growth was seen on each day with signs of maturation evident by day 7 in culture. The polyester-based filter inserts can be substituted in the previously established permeability assay without alteration of morphology or rate of growth, thus allowing confirmation of cell confluency prior to use in the permeability assay. It will also allow for photographic documentation of cell injury prior to recovery studies, without necessitating the preparation of extra samples for fixation at the time of initial injury. Future studies will be required to determine if there will be any alteration in the rate of permeability with a previously tested standard, before adoption of the new filter.

show abstract

Section: Discussionsupporting

confidence: 65%

Section: Introductionmentioning

confidence: 98%

Comparative Development of SIRC Rabbit Corneal Cells Grown on Polycarbonate- and Polyester-Based Filters

Hutak

Kavanagh

Reddy

2002

Skin Pharmacol Physiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Some investigators refer to the cell line as fibroblastic, 24 whereas others claim that it is of epithelial origin. [25][26][27] Earlier reports from our laboratory have shown that these cells indeed display epithelial morphology. 31 Several studies reported good correlation between permeability coefficients across SIRC cell line and intact rabbit cornea.…”

Section: Introductionmentioning

confidence: 99%

“…The SIRC cell line was originally derived from the cornea of a normal hare 24 and has been widely used as an in vitro model to assess corneal transport as well as immunological and toxicological responses. [25][26][27][28][29][30][31] However, contradictory reports exist about the epithelial origin of the cell line. Some investigators refer to the cell line as fibroblastic, 24 whereas others claim that it is of epithelial origin.…”

Section: Introductionmentioning

confidence: 99%

Functional differences in nucleoside and nucleobase transporters expressed on the rabbit corneal epithelial cell line (SIRC) and isolated rabbit cornea

et al. 2003

View full text Add to dashboard Cite

ABSTRACTclovir permeates the cornea primarily by passive diffusion. Nucleoside and nucleobase transporters on rabbit cornea and corneal epithelial cell line, SIRC, are functionally different, undermining the utility of the SIRC cell line as an in vitro screening tool for elucidating the corneal permeation mechanism of nucleoside analogs. Uptake and transport studies were performed for 2 minutes and 120 minutes, respectively, at 34°C. Thymidine uptake by SIRC displayed saturable kinetics (K m = 595.9 ± 80.4μM, and V max = 289.5 ± 17.2 pmol/min/mg protein). Uptake was inhibited by both purine and pyrimidine nucleosides but not by nucleobases. [ 3 H]thymidine uptake was sodium and energy independent but was inhibited by nitrobenzylthioinosine at nanomolar concentrations. Adenine uptake by SIRC consisted of a saturable component (K m = 14.4 ± 2.3μM, V max = 0.4 ± 0.04 nmol/min/mg protein) and a nonsaturable component. Uptake of adenine was inhibited by purine nucleobases but not by the nucleosides or pyrimidine nucleobases and was independent of sodium, energy, and nitrobenzylthioinosine. [ 3 H]ganciclovir uptake involved a carrier-mediated component and was inhibited by the purine nucleobases but not by the nucleosides or pyrimidine nucleobases. However, transport of [ 3 H]adenine across the isolated rabbit cornea was not inhibited by unlabeled adenine. Further, corneal permeability of ganciclovir across a 100-fold concentration range remained constant, indicating that ganci-

show abstract

“…As recent works [8] showed that epithelial cells cultured in air-liquid interface conditions grow mostly in a multilayer fashion, we chose two different approaches, growing the cells on a collagen-coated permeable support under a liquid-covered condition (LCC) and under an air interface condition (AIC). As part of the characterization of the BCEC layers, we determined the culture's responses to toxic treatments.…”

Section: Introductionmentioning

confidence: 99%

Multilayer Primary Epithelial Cell Culture from Bovine Conjunctiva as a Model for in vitro Toxicity Tests

Civiale¹,

Paladino²,

Marino³

et al. 2003

Ophthalmic Res

View full text Add to dashboard Cite

Objective: The purpose of this study was to obtain a primary cell culture of bovine origin similar to the conjunctiva in terms of morphology and cell types, which could be of use for in vitro toxicity studies. Methods: After separation from the stroma by enzymatic treatment, conjunctival epithelial cells were dissociated and plated onto collagen-coated Transwell^® filters (1.13 cm² area). One group of plates was maintained in immersion and another was cultured under air-lifted conditions. Anti-epithelial keratin antibodies (AE1/AE3, K4) and antidesmoplakin 1 and 2 were used to characterize the cells by indirect immunofluorescence. The cell layer was examined after histological processing of the Transwell filter. Ultrastructural analysis was carried out by scanning electron microscopy (SEM). The bioelectric parameters transepithelial electrical resistance (TEER), potential difference (PD), short circuit current and paracellular permeability profile of carboxyfluorescein were monitored as indices of the functional characteristics of these cultures. Cytotoxicity was evaluated on morphological and functional (TEER) grounds after treating the cultures with several test substances. Results: Morphological studies showed pure and homogeneous cell cultures. In the SEM analysis, we observed contiguous polygonal cells with numerous short microvilli, a characteristic proportion of light, medium and dark cells and a sparse population of rounded PAS-positive cells, i.e. resembling goblet cells. Air-lifted cultures also showed a tissue-like cellular organization (8–9 layers). Immersion cultures reached a maximum TEER value of around 2.95 kΩ·cm² 7 days after plating while in air-lifted cultures TEER peaked up to 5.59 kΩ·cm² 11 days after plating. With regard to the use of bovine conjunctival epithelial cells (BCECs) for cytotoxicity screening, the system responded finely to the insults and yielded morphological and functional results in accordance with data obtained in vivo. Conclusions: BCECs reproduce cell morphology and differentiation of the original tissue and should prove a useful tool for initial studies of drug toxicity.

show abstract

Growth Pattern of Sirc Rabbit Corneal Cells in Microwell Inserts

Cited by 16 publications

References 9 publications

Comparative Development of SIRC Rabbit Corneal Cells Grown on Polycarbonate- and Polyester-Based Filters

Comparative Development of SIRC Rabbit Corneal Cells Grown on Polycarbonate- and Polyester-Based Filters

Functional differences in nucleoside and nucleobase transporters expressed on the rabbit corneal epithelial cell line (SIRC) and isolated rabbit cornea

Multilayer Primary Epithelial Cell Culture from Bovine Conjunctiva as a Model for in vitro Toxicity Tests

Contact Info

Product

Resources

About