Charbonneau, A.M., Al‐Samadi, A., Salo, T. and Tran, S.D. (2019), 3D Culture Histology Cryosectioned Well Insert Technology Preserves the Structural Relationship between Cells and Biomaterials for Time‐Lapse Analysis of 3D Cultures. Biotechnol. J., 14: 1900105. doi:10.1002/biot.201900105
3D culture histology cryosectioned well insert technology preserves the structural relationship between cells and biomaterials for time‐lapse analysis of 3D cultures
|Author:||Charbonneau, André M.1; Al‐Samadi, Ahmed2; Salo, Tuula2;|
1Faculty of Dentistry, McGill University, 3640 University Street, H3A 0C7, Montréal, Canada
2Oral and Maxillofacial Diseases, University of Helsinki, Helsinki, Finland
|Online Access:||PDF Full Text (PDF, 16.6 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe202001172512
John Wiley & Sons,
|Publish Date:|| 2020-07-11
When performing histology of softer biomaterials, aspiration disrupts the cellular and molecular location information. This study aims to develop a cryosectionable well insert able to preserve the biomaterial and cell’s original 3D conformation from the well to histology analysis. The well insert is composed of a paraffin‐coated gelatine pill. Within the coated capsule, the human epithelial cell line (NS‐SV‐AC) is cultured in Matrigel, GrowDex, Myogel, Myogel + GrowDex, or cell culture media for 14 days. At 0 and 14 days, the samples are frozen in liquid nitrogen and cryotome is used to create sections. The slides are stained by Sirius Red and immunohistochemistry using antibodies human collagens I–V and human Ki‐67. Sirius Red shows pink shades of biomaterials and the best cellular vertical distribution throughout the sagittal section of the well is achieved with Matrigel, GrowDex, and Myogel + GrowDex; in Myogel and media, the cells sink. For collagen protein expression, only Matrigel induces a notable difference while in the other materials, collagen staining is weak or difficult to distinguish from endogenous collagens. Ki‐67 expression is maintained over time. The 3D‐cryo well insert provides a new time‐lapse histology perspective of analysis for liquid or gel cultures that maintains cells and macromolecules in their unaltered in‐well configuration.
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
The authors thank Canadian funding agencies MITACS (Globablink) and FRSQ (Doctoral Training) for their research funds for graduate students.
© 2019 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. This is the peer reviewed version of the following article: Charbonneau, A.M., Al‐Samadi, A., Salo, T. and Tran, S.D. (2019), 3D Culture Histology Cryosectioned Well Insert Technology Preserves the Structural Relationship between Cells and Biomaterials for Time‐Lapse Analysis of 3D Cultures. Biotechnol. J., 14: 1900105, which has been published in final form at https://doi.org/10.1002/biot.201900105. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving