Journal: Advanced Science

Article Title: The Bioengineering of Microspheric Skin Organoids and Their Application in Drug Screening

doi: 10.1002/advs.202416863

Figure Lengend Snippet: Fabrication of microspheric skin organoids. A). A schematic diagram (Created in BioRender) illustrating the formation of microspheric skin organoids. B). Photographs of human dermal fibroblast (HDF)‐spheres at day 0 and day 1 under a light microscopy. C). Representative images of day 5 microspheric skin organoids (left), and the cross section of the organoid after immunofluorescent staining for E‐cadherin (E‐cad) and vimentin. D). Representative images of cross‐sections of a day 7 organoid after immunofluorescence staining for F‐actin and E‐cad. E). Representative microscopic images of a day 7 microspheric skin organoid (whole mount) showing a core‐shell structure, with the core composed of mCherry‐expressing HDFs (red) and the shell formed by GFP‐expressing HaCaT cells (green). Scale bar: 100 µm.

Article Snippet: HaCaT cells, human foreskin primary epidermal stem cells (Epi‐SCs), and human dermal fibroblasts (HDFs) were purchased from iCell Bioscience Co., Ltd (Shanghai).

Techniques: Light Microscopy, Staining, Immunofluorescence, Expressing

Journal: Advanced Science

Article Title: The Bioengineering of Microspheric Skin Organoids and Their Application in Drug Screening

doi: 10.1002/advs.202416863

Figure Lengend Snippet: Terminal differentiation of the epidermis in microspheric skin organoids. A). A schematic diagram (Created in BioRender) illustrating the formation of cornified microspheric skin organoids. Organoids consisting of primary human keratinocytes and HDFs were first cultured in growth medium to allow keratincoytes to proliferate, and then in induction medium to induce epidermal maturation. B). Representative images of microspheric skin organoids prepared by different keratinocytes. Cytokeratins were stained using a pan‐cytokeratin (Pan‐CK) antibody, E‐cadherin (E‐cad) was stained in red, and nuclei were stained blue. C). Lucifer Yellow permeability assay of the human dermal fibroblast core (HDF, left), skin organoids with epidermis formed by HaCaT cells (middle), or by primary human keratinocytes (right), where the dye was in green color and the nuclei were in blue. D). Sections of day 7 organoids comprising primary human keratinocytes were stained for keratin 5, keratin 10, and filaggrin to illustrate the differentiation of the epidermal cells. Scale bars: 100 µm.

Article Snippet: HaCaT cells, human foreskin primary epidermal stem cells (Epi‐SCs), and human dermal fibroblasts (HDFs) were purchased from iCell Bioscience Co., Ltd (Shanghai).

Techniques: Cell Culture, Staining, Permeability