Changes in keratins and alpha-smooth muscle actin during three-dimensional reconstitution of eccrine sweat glands

Abstract

We have examined the changes of keratins and alpha-SMA at various time points in order to investigate the development and differentiation of eccrine sweat gland cells during the course of three-dimensional (3D) reconstitution. Mixtures of eccrine sweat gland cells and Matrigel were injected subcutaneously into the inguinal regions of nude mice. At 1, 2, 4, 6, 8, 14, 21, 28, 35, and 42 days post-implantation, Matrigel plugs were removed and immunostained. We found that during 3D reconstitution, keratin and alpha-SMA expression changed in a time-dependent manner. At day 1, all cells stained positively for keratin isoforms K5, K14, and K15, with the staining intensity of K15 being weak and K5 and K14 being strong, but none of the cells displayed K7, K8, or alpha-SMA. As time progressed, spheroid-like structures formed with the inner layer acquiring K7 and K8, but losing K5 and K14 expression, and the outer layer acquiring alpha-SMA expression, but losing K15 expression. K8 expression was first noted at day 14, and K7 and alpha-SMA at day 21. The loss of K15 expression was first noted at day 14, K14 at day 21, and K5 at day 28. At 28, 35, and 42 days, the spheroid-like structures could be distinguished, by immunohistochemistry, as having secretory coil-like and coiled duct-like structures. We conclude that the changes in expression of keratins and alpha-SMA in 3D-reconstituted eccrine sweat glands are similar to those of native eccrine sweat glands, indicating that the 3D reconstitution of sweat glands provides an excellent model for studying the development, cytodifferentiation, and regulation of eccrine sweat glands.

A novel cell line from human eccrine sweat gland duct cells for investigating sweating physiology

“…This suggested a certain self-organization potential of isolated eSG cells with some degree of cell-cell communication and interaction in those scaffold-free 3D HD models. Self-organization and -differentiation of coil and duct cells in in vitro models were also reported by other research groups using Matrigel-assisted eSG replicas and their implantation into nude mice [69,70].…”

A novel organotypic 3D sweat gland model with physiological functionality

“…Second, the epithelial apical-basal polarity within the 3D model was shown by histochemical analysis with specific SG cell marker proteins like AQP5, CHRM3, CEACAM5, and ANO1. Li et al demonstrated a comparable reconstruction of sweat gland-like structures with Matrigel-cultivated primary cells, which only formed tubular-like structures after transplantation in mice [ 36 , 43 ]. However, the transplantation step makes their model more complicated as compared to our hanging-drop culture and would not be suitable for applications, which prohibit the use of animal models such as screening for cosmetic bioactives.…”

An innovative application of follicular unit extraction technique in the treatment of bromhidrosis

“…The decapitation products were positive for K5, Brst-2 and CEA, the commonly used markers for both apocrine and eccrine glands. 14,15 It was worth noting that no obvious inflammatory reaction was observed, which demonstrated that the interaction between apocrine gland secretions and bacteria did not occur in the body.…”