Xie Sitian, Chen Liyun, Zhang Mingjun, Zhang Cuiping, Li Haihong
Department of Plastic Surgery and Burn Center, The Second Affiliated Hospital, Shantou University Medical College, Shantou, China.
Wound Healing and Cell Biology Laboratory, The First Affiliated Hospital, Chinese PLA General Hospital, Beijing, China.
Ann Transl Med. 2022 Jul;10(14):767. doi: 10.21037/atm-22-3252.
3D organoid cultures of hair follicles (HFs) are powerful models that mimic native HF for both in-depth study of HF disease and precision therapy. However, few studies have investigated the complete structure and properties of HF organoids. To investigate and characterize the complete HF organoids self-assembled by coculture of neonatal mouse epidermal cells (MECs) and dermal cells in Matrigel.
Fresh epidermal and dermal cells from newborn mice (n=4) were isolated, and cocultured (1:1 ratio) in Matrigel using DMEM/F12 medium for 1 week. During the culture, an inverted microscope was used to observe the morphology of the 3D constructs. After 1 week, hematoxylin-eosin (HE) and immunofluorescence (IF) staining of HF-related markers (K5, K73, AE13, and K10), HF stem cell markers (K15, CD34, CD49f), skin-derived precursor-related marker (Nestin), and dermal papillae (DP)-specific markers (SOX2 and ALP) was performed in the harvested constructs to identify the HF organoids.
Epidermal and dermal cells self-assembled into HF organoids comprising an infundibular cyst-like structure, a lower segment-like structure, and a bulb-like structure from tail to root. The HF organoid had multiple, well-defined compartments similar to native anagen HF. Of the three segments, K73 was expressed in the inner root sheath-like layer, AE13 was localized in the hair shaft-like structure, K5, K15, CD34, and CD49f were present in the outer root sheath-like layer, Nestin labeled the connective tissue sheath-like layer, and SOX2 and ALP were expressed in the DP-like structure. Furthermore, K10 and K73 were expressed in the infundibular cyst-like structure. The expression of these molecular proteins was consistent with native anagen HF.
The complete HF organoid regenerated in Matrigel has specific compartments and is an excellent model to study HF disease and precision therapy.
毛囊的3D类器官培养是强大的模型，可模拟天然毛囊用于深入研究毛囊疾病和精准治疗。然而，很少有研究调查毛囊类器官的完整结构和特性。为了研究和表征通过在基质胶中共同培养新生小鼠表皮细胞（MECs）和真皮细胞自组装而成的完整毛囊类器官。
从新生小鼠（n = 4）中分离新鲜的表皮和真皮细胞，并使用DMEM/F12培养基在基质胶中以1:1的比例共同培养1周。在培养过程中，使用倒置显微镜观察3D构建体的形态。1周后，对收获的构建体进行苏木精-伊红（HE）和免疫荧光（IF）染色，检测毛囊相关标志物（K5、K73、AE13和K10）、毛囊干细胞标志物（K15、CD34、CD49f）、皮肤衍生前体相关标志物（巢蛋白）和毛乳头（DP）特异性标志物（SOX2和碱性磷酸酶），以鉴定毛囊类器官。
表皮和真皮细胞自组装成毛囊类器官，从尾端到根部包括漏斗状囊肿样结构、下段样结构和球状结构。毛囊类器官具有多个界限分明的隔室，类似于天然生长期毛囊。在这三个节段中，K73在内根鞘样层表达，AE13定位于毛干样结构中，K5、K15、CD34和CD49f在外根鞘样层中存在，巢蛋白标记结缔组织鞘样层，SOX2和碱性磷酸酶在DP样结构中表达。此外，K10和K73在漏斗状囊肿样结构中表达。这些分子蛋白的表达与天然生长期毛囊一致。
在基质胶中再生的完整毛囊类器官具有特定的隔室，是研究毛囊疾病和精准治疗的优秀模型。
