Development of human ectocervical tissue models with physiologic endocrine and paracrine signaling†

Abstract

<jats:title>Abstract</jats:title> <jats:p>There is a shortage of research models that adequately represent the unique mucosal environment of human ectocervix, limiting development of new therapies for treating infertility, infection, or cancer. We developed three microphysiologic human ectocervix models to study hormone action during homeostasis. First, we reconstructed ectocervix using decellularized extracellular matrix scaffolds, which supported cell integration and could be clinically useful. Secondly, we generated organotypic systems consisting of ectocervical explants co-cultured with murine ovaries or cycling exogenous hormones, which mimicked human menstrual cycles. Finally, we engineered ectocervix tissue consisting of tissue-specific stromal-equivalents and fully-differentiated epithelium that mimicked in vivo physiology, including squamous maturation, hormone response, and mucin production, and remained viable for 28 days in vitro. The localization of differentiation-dependent mucins in native and engineered tissue was identified for the first time, which will allow increased efficiency in mucin targeting for drug delivery. In summary, we developed and characterized three microphysiologic human ectocervical tissue models that will be useful for a variety of research applications, including preventative and therapeutic treatments, drug and toxicology studies, and fundamental research on hormone action in a historically understudied tissue that is critical for women’s health.</jats:p>

Characterization of 3D organotypic epithelial tissues reveals tonsil-specific differences in tonic interferon signaling

“…The estrogen receptor ( ESR1) was up-regulated in tissues with a genital origin. Estrogen receptors are highly expressed in the ectocervix (McKinnon et al, 2020) and foreskin (Pask et al, 2008), and we found it was highest in cervix (Wald P -adj = 5.82 x 10 -7 and 9.89 x 10 -20 , relative to foreskin and tonsil, respectively), followed by foreskin (Wald P -adj = 1.40 x 10 -3 relative to tonsil), and lowest in tonsil ( Figure 7H ).…”

“…While modifications can be made to these 3D models depending on the experimental context and complexity desired, we used an organotypic 3D raft model with the same fibroblasts, collagen, media, growth conditions, and duration to maximize our ability to compare between tissues. For example, 3D organotypic skin and cervical models have been embedded with additional cell types, such as Langerhans cells (Jackson et al, 2020a;Kosten et al, 2015), and others have tested tissue-specific hormone supplementation (estrogen and progesterone) when culturing 3D organotypic ectocervical epithelium (McKinnon et al, 2020). A primary goal of our study was to provide evidence that these different in vitro tissues maintain similarities with their in vivo tissue equivalents, independent of other cell types or microbiota normally present in these tissues.…”

Future Challenges and Opportunities of Extracellular Matrix Hydrogels in Female Reproductive Medicine

“…ECM products that are obtained from decellularization of female reproductive organs have salient roles in the field of reproductive medicine, with research focusing mainly on those that are derived from the uterus [ 51 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 ], ovary [ 50 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 ], cervix [ 93 ], vagina [ 94 , 95 ], and placenta [ 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 ]. The variable methods that are used for decellularization are dependent on the tissue type and its characteristics.…”

In vitro modelling of the physiological and diseased female reproductive system

“…To further enhance the biomimetic aspects of the FRT in vitro models, they can contain tissue-specific ECM [171]. This can be achieved by either using decellularized tissue-specific ECM [172,173] or by making use of the ability of cells to secrete ECM proteins around themselves in vitro [174], both of which are methods that provide a more physiological ECM ligand milieu compared to traditionally used collagen and Matrigel. While there are several chemical and physical decellularization techniques, all of these techniques aim to remove the cellular component and cellular antigens while preserving the ECM proteins and their mechanical properties [175].…”

“…• Collagen I [238,260,261] • Matrigel [68] • Decellularized ECM [172,173,178,262] • Chemically Crosslinked Collagen [238] • Alginate [263] • PEG [264][265][266] • Silk fibroin [267,268] • GeLMA [269] • Peptide amphiphiles [209] • PLGA-PEG-PLGA [270] • Fibrin-based matrix [142,143] • Tyramine-based Hyaluronan (HA) Hydrogel [271] • Potential for dynamic crosslinks and on-demand ligand and stiffness modulation Identifying the materialindependent macroscale scaffold properties (e.g. porosity) on functional readouts Materials that support different cell types…”