Human stroma and epithelium co-culture in a microfluidic model of a human prostate gland

Abstract

The prostate is a walnut-sized gland that surrounds the urethra of males at the base of the bladder comprising a muscular portion, which controls the release of urine, and a glandular portion, which secretes fluids that nourish and protect sperms. Here, we report the development of a microfluidic-based model of a human prostate gland. The polydimethylsiloxane (PDMS) microfluidic device, consisting of two stacked microchannels separated by a polyester porous membrane, enables long-term in vitro cocultivation of human epithelial and stromal cells. The porous separation membrane provides an anchoring scaffold for long-term culturing of the two cell types on its opposite surfaces allowing paracrine signaling but not cell crossing between the two channels. The microfluidic device is transparent enabling high resolution bright-field and fluorescence imaging. Within this coculture model of a human epithelium/stroma interface, we simulated the functional development of the in vivo human prostate gland. We observed the successful differentiation of basal epithelial cells into luminal secretory cells determined biochemically by immunostaining with known differentiation biomarkers, particularly androgen receptor expression. We also observed morphological changes where glandlike mounds appeared with relatively empty centers reminiscent of prostatic glandular acini structures. This prostate-on-a-chip will facilitate the direct evaluation of paracrine and endocrine cross talk between these two cell types as well as studies associated with normal vs disease-related events such as prostate cancer.

The Role of Cancer-Associated Fibroblasts in Prostate Cancer Tumorigenesis

“…In this section, we discuss the multiple ways in which CAFs affect the primary PCa microenvironment and tumor growth, particularly regarding ECM modulation, signaling interaction mechanisms with tumor cells and androgen regulation. Different methods have been implemented for the study of prostate epithelial-stromal interactions, such as the in vivo smooth muscle invasion xenograft model to assess the invasive potential of human tumor cells into the smooth muscle/skeletal muscle structures of the diaphragm [112], the differential reactive stroma xenograft model [107], the 3D co-culture of organoids with prostate stromal cells [113], and the novel microfluidic human prostate in a chip ex vivo model [114].…”

Comprehensive Development in Organ-On-A-Chip Technology

“…The development of the OOC platform can revolutionaries the drug development process. Physiologically relevant mimicking of the microenvironment and ability to incorporate physiological forces affecting niche and co-culturing of multiple types of the cell and Jiang et al 15 Kidney-on-a-chip For predicting the renal tubular reabsorption of various drugs Primary human renal proximal tubule epithelial cells (RPTECs), Sakolish et al 16 Endometrium-on-a-chip For studying the role of hemodynamic forces on endometrial decidualization…”

Engineering cell–substrate interactions on porous membranes for microphysiological systems

“…29 Further evidence is presented by Kim et al where they suggested a deterministic role for TAZ regulation in upregulation of PDX1+ and enhancement of pancreatic differentiation (Table 1). 47 Due to the increasing interest in guiding stem cell fate over porous membranes in co-culture systems, [49][50][51] the role of substrate disruption in cell differentiation should be considered in these studies. Moreover, these findings can also be employed to help develop more physiologically relevant cellular responses and direct cell phenotype by optimizing the design of porous membranes.…”

“…Due to the increasing interest in guiding stem cell fate over porous membranes in co-culture systems, 49–51 the role of substrate disruption in cell differentiation should be considered in these studies. Moreover, these findings can also be employed to help develop more physiologically relevant cellular responses and direct cell phenotype by optimizing the design of porous membranes.…”