Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids

Abstract

Summary The mammalian hair follicle arises during embryonic development from coordinated interactions between the epidermis and dermis. It is currently unclear how to recapitulate hair follicle induction in pluripotent stem cell cultures for use in basic research studies or in vitro drug testing. To date, generation of hair follicles in vitro has only been possible using primary cells isolated from embryonic skin, cultured alone or in a co-culture with stem cell-derived cells, combined with in vivo transplantation. Here, we describe the derivation of skin organoids, constituting epidermal and dermal layers, from a homogeneous population of mouse pluripotent stem cells in a 3D culture. We show that skin organoids spontaneously produce de novo hair follicles in a process that mimics normal embryonic hair folliculogenesis. This in vitro model of skin development will be useful for studying mechanisms of hair follicle induction, evaluating hair growth or inhibitory drugs, and modeling skin diseases.

Hair-bearing human skin generated entirely from pluripotent stem cells

“…NOTE: In both foetal and xenograft HFs, NFATC1 expression is predominantly localized to the cytoplasm in bulge cells. By contrast, adult facial HFs have bulge regions have cells with nuclear-localized NFATC1—reminiscent of previous reports showing nuclear-localized NFATC1 in mouse bulge stem cells 4,38 . Dashed boxes indicate magnified regions shown on a corner of each image panel (a, c, d).…”

“…Whole-mount staining was performed with a previously published method 43 with major adjustments in incubation times to account for reduced tissue size 4 . In brief, samples were fixed in 4% (v/v) PFA (Electron Microscopy Sciences), permeabilized, descaled, incubated with primary antibodies, incubated with fluorescently-labelled secondary antibodies, re-fixed in 4% (v/v) PFA (Electron Microscopy Sciences), and cleared.…”

“…For decades, culture systems containing epidermal and dermal cells have been used to model human skin outside of the body 3,9,10 ; however, it has been persistently difficult to grow and maintain functional skin appendages in culture. We recently reported that appendage-bearing skin organoids can be generated via directed differentiation of epidermal and dermal cells from mouse pluripotent stem cells (PSCs) 4 . From this study, we recognised that a 3D cyst composed of surface ectoderm (i.e.…”

Organoids – New Models for Host–Helminth Interactions

“…Organoid systems were initially developed to recreate murine and human small intestine and colon epithelia, and involved the use of specific culture media cocktails to recreate the SCN conditions for each intestinal segment in the specific species [10,11,17]. Subsequently, organoids have been developed for a great variety of tissues and organs, including stomach, esophagus, liver, lung, pancreas, prostate, brain, kidney, mammary gland, ovary, lingual, taste bud, salivary gland, testis, endometrium, Fallopian tube, lymph nodes, blood vessels, skin, inner ear, and retina [1,2,11,[18][19][20][21]. Moreover, intestinal, mammary, keratinocyte, and liver organoids from other animal models already exist, including those of bovine, porcine, ovine, chicken, feline, and canine origin [22].…”

Comparative models for human nasal infections and immunity

“…The first organoid was created 9 years ago from a single intestinal crypt cell; where even in the absence of a non-epithelial cellular niche, a single cell was able to self-renew and differentiate, generating a villus-like epithelial structure in which all the different cell types present in the real organ were found (Sato et al, 2009). Since then several "mini-tissue" have been reproduced successfully in many animals including human pancreas, kidney, thyroid, liver, inner ear, retina, skin tissue with follicular hair and brain (Barkauskas et al, 2017;Lee et al, 2018;Koehler et al, 2017;Orsini et al, 2018;Ramachandran et al, 2015;Turksen, 2016;Ueda et al, 2018).…”