Novel Three Dimensional Human Endocervix Cultures Respond to 28-Day Hormone Treatment

Abstract

The endocervix has both anatomical and biological functions that participate in the delicate balance between tolerance necessary for conception and protection from pathogens. Our goal was to develop a robust 3-dimensional (3D) endocervix model that was a reliable representation of the in vivo tissues and to identify the physiological responses to changing levels of steroid hormones during a 28-day time period. Human endocervical cells were grown on polystyrene scaffolds, and the morphologic and hormonal responses of cultured cells were assessed in response to fluctuating levels of estradiol (E2) or progesterone (P4). Morphologically, the 3D cultures were composed of a mixed population of cells, including epithelial and stromal cells. Treatment with E2 and P4 (d 28) increased cell growth and proliferation as compared with no treatment control. Cells expressed estrogen receptor and P4 receptor and produced both neutral and acidic mucins, including Mucin 16. In addition, a 45-plex Luminex assay identified numerous factors secreted and regulated by hormones. Specifically, IL-1β and leukemia inhibitory factor significantly decreased in the presence of E2 and P4 as compared with the no hormone control at day 26. Cotreatment with RU486 (mifepristone) attenuated the inhibition of IL-1β and leukemia inhibitory factor secretion. In summary, a robust, novel 3D endocervical culture was developed, and physiologic responses to the menstrual cycle mimic of E2 and P4 levels for a period of 28 days were identified.

Distinct Roles of Cervical Epithelia and Stroma in Pregnancy and Parturition

“…hormonally regulated endocervical epithelial culture system that provides a useful system to understand the barrier and immunomodulatory roles of the cervical epithelia has recently been described. 53 Another potentially valuable tool is the ongoing development of mathematical models that allow the integration of dynamic changes in the hormone environment, ECM composition and structure, and mechanical properties of the cervix. 54 Such mathematical frameworks provide insights into structure-function relationships.…”

Differentiation Patterns of Immortalized Human Meibomian Gland Epithelial Cells in Three-Dimensional Culture

“…37,38 Histologic analysis demonstrated that HMGECs exhibited the same appearance regardless of their location within Alvetex. We expected that air-liquid interface culture encouraged the epithelial stratification upon Alvetex; however, HMGECs did not form multiple cell layers, but rather differentiated into sphere-shaped colonies on the apical surface as already observed in membrane-based 3D air-lift culture.…”

Development of a novel human recellularized endometrium that responds to a 28-day hormone treatment†

“…The concentration of hormones used in our 28-day protocol was based on clinical data of serum levels of hormones in women during the menstrual cycle [53,54]. Our group has previously used this treatment protocol on endocervical cells grown in a 3D polystyrene (Alvatex) scaffold [29] and primary fallopian tube explants [55] showing viability and hormone response in both tissues. One of the hallmarks of hormone response of endometrial cells is the decidualization response that becomes evident during the late secretory phase in endometrial stromal cells surrounding the spiral arteries and involves their transformation into differentiated, secretory decidual cells [56].…”

“…A stepwise hormone protocol was used to treat cells within the recellularized scaffolds to mimic the human 28-day menstrual cycle as previously described with some modifications [29]. Recellularized scaffolds were divided into three treatment groups.…”