title: Surface Tension Guided Hanging-Drop: Producing Controllable 3D Spheroid of High-Passaged Human Dermal Papilla Cells and Forming Inductive Microtissues for Hair-Follicle Regeneration.
abstract: Human dermal papilla (DP) cells have been studied extensively when grown in the conventional monolayer. However, because of great deviation from the real in vivo three-dimensional (3D) environment, these two-dimensional (2D) grown cells tend to lose the hair-inducible capability during passaging. Hence, these 2D caused concerns have motivated the development of novel 3D culture techniques to produce cellular microtissues with suitable mimics. The hanging-drop approach is based on surface tension-based technique and the interaction between surface tension and gravity field that makes a convergence of liquid drops. This study used this technique in a converged drop to form cellular spheroids of dermal papilla cells. It leads to a controllable 3Dspheroid model for scalable fabrication of inductive DP microtissues. The optimal conditions for culturing high-passaged (P8) DP spheroids were determined first. Then, the morphological, histological and functional studies were performed. In addition, expressions of hair-inductive markers including alkaline phosphatase, α-smooth muscle actin and neural cell adhesion molecule were also analyzed by quantitative RT-PCR, immunostaining and immunoblotting. Finally, P8-DP microtissues were coimplanted with newborn mouse epidermal cells (EPCs) into nude mice. Our results indicated that the formation of 3D microtissues not only endowed P8-DP microtissues many similarities to primary DP, but also confer these microtissues an enhanced ability to induce hair-follicle (HF) neogenesis in vivo. This model provides a potential to elucidate the native biology of human DP, and also shows the promising for the controllable and scalable production of inductive DP cells applied in future follicle regeneration.
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.|Department of Mechanical Engineering and Department of Biomedical & Medical Genetics, University of Manitoba , 75A Chancellors Circle, Winnipeg, Manitoba R3T 2N2, Canada.|Children's Hospital Research Institute of Manitoba , 715 McDermot Avenue, Winnipeg, Manitoba R3E 3P4, Canada.
authors.forename: Bojie
authors.initials: B
authors.lastname: Lin
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Miao
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Zhexiang
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Du
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Su
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Bingcheng
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University , 1838 North Guangzhou Avenue, Guangzhou, Guangdong 510515, China.
authors.forename: Zhiqi
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Department of Mechanical Engineering and Department of Biomedical & Medical Genetics, University of Manitoba , 75A Chancellors Circle, Winnipeg, Manitoba R3T 2N2, Canada.|Children's Hospital Research Institute of Manitoba , 715 McDermot Avenue, Winnipeg, Manitoba R3E 3P4, Canada.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Xing
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D020405:Dermis / Q000166:cytology* / Q000378:metabolism; D018859:Hair Follicle / Q000473:pathology / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D019006:Neural Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012038:Regeneration / Q000502:physiology*; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism; D013500:Surface Tension
chemical_list: D019006:Neural Cell Adhesion Molecules
keywords: 3D culture; dermal papilla; hair-follicle induction; hanging-drop; surface tension