Establishment and optimization of epithelial cell cultures from human ectocervix, transformation zone, and endocervix optimization of epithelial cell cultures

• Published in: Journal of cellular physiology 2019-06, Vol.234 (6), p.7683-7694
• Main Authors: Deng, Han, Mondal, Sumona, Sur, Shantanu, Woodworth, Craig D.
• Format: Article
• Language: English
• Subjects: Biotechnology; Carcinogenesis; Carcinogenesis - pathology; Carcinogens; Cell adhesion; Cell culture; Cell Culture Techniques - methods; Cell Differentiation - physiology; Cervical cancer; Cervix Uteri - cytology; Collagen; Culture media; Differentiation (biology); ectocervix; endocervix; Epithelial cells; Epithelial Cells - cytology; Epithelium; Epithelium - pathology; Female; human papillomavirus (HPV); Humans; Mucous membrane; Optimization; organotypic culture; Public health; Rafts; Stromal cells; transformation zone (TZ); Uterine Cervical Neoplasms - pathology
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.28049

Cervical cancer is a major public health problem and research using cell culture models has improved understanding of this disease. The human cervix contains three anatomic regions; ectocervix with stratified squamous epithelium, endocervix with secretory epithelium, and transformation zone (TZ) with metaplastic cells. Most cervical cancers originate within the TZ. However, little is known about the biology of TZ cells or why they are highly susceptible to carcinogenesis. The goal of this study was to develop and optimize methods to compare growth and differentiation of cells cultured from ectocervix, TZ or endocervix. We examined the effects of different serum‐free media on cell attachment, cell growth and differentiation, and cell population doublings in monolayer culture. We also optimized conditions for organotypic culture of cervical epithelial cells using collagen rafts with human cervical stromal cells. Finally, we present a step‐by‐step protocol for culturing cells from each region of human cervix. The goal of this study was to develop and optimize methods to compare growth and differentiation of cells cultured from ectocervix, transformation zone (TZ) or endocervix. We examined the effects of different serum‐free media on cell attachment, cell growth and differentiation, and cell population doublings in monolayer culture. In addition, we present a step‐by‐step protocol for culturing cells from each region of human cervix.