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

Abstract

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.

Primary ectocervical epithelial cells display lower permissivity to Chlamydia trachomatis than HeLa cells and a globally higher pro-inflammatory profile

“…Compared to other methods in which enzymatic cocktails were applied to isolate primary epithelial cells from the fallopian tubes and the uterus of human samples 14 , 26 , 27 , a single enzyme Dispase ® II was used. A similar methodology was used by other groups in isolating primary cervical epithelial cells 21 , 28 . One important step we introduced was the mechanical separation of the mucosal layer from the stromal tissue before dissociating the epithelial cells, which significantly improved the purity of isolated cells.…”

KCa3.1-dependent uptake of the cytotoxic DNA-binding dye Hoechst 33258 into cancerous but not healthy cervical cells

“…Cervical carcinoma cell lines (CXT) and nonmalignant cervical epithelial cell strains (HCX) were derived from tissues purchased from the Cooperative Human Tissue Network as described elsewhere ( 50 ). The samples were surgically removed for other purposes, and patient origin was not identified.…”

Biomaterial strategies to replicate gynecological tissue

“…174 A separate study used 3D organotypic cultures with human stromal and epithelial cells from each cervical region to evaluate the invasion of epithelial cells and their interaction with stromal cells. 109 Increasing the concentration of stromal cells increased the invasion of epithelial cells from each cervical region, suggesting that stromal-epithelial interactions play an essential role in cervical tissue homeostasis. 109 The establishment of organotypic raft cultures of cervical cells represents a crucial step forward in understanding cervix biology and function.…”

“…109 Increasing the concentration of stromal cells increased the invasion of epithelial cells from each cervical region, suggesting that stromal-epithelial interactions play an essential role in cervical tissue homeostasis. 109 The establishment of organotypic raft cultures of cervical cells represents a crucial step forward in understanding cervix biology and function.…”