细胞技术

In this article we provide a method to isolate hair follicle stem cells that have undergone targeted gene inactivation. The mice from which these cells are isolated are bred into a Rosa26-yellow fluorescent protein (YFP) reporter background, which results in YFP expression in the targeted st ...

Skin regeneration is intricately controlled by epidermal stem cells. In human skin, the long-lived, slow-cycling, and highly proliferative stem cells are located in the basal layer of the interfollicular epidermis (IFE). The ability to isolate and culture human IFE stem cells (IFESCs) of ...

The skin stem cells are located at two distinct locations, the basement of epidermis and the hair follicle bulge. The bulge stem cell is considered to be of higher hierarchy in terms of the stemness than the epidermal stem cell. Recently, hair follicle bulge cells can be successfully isolated using fl ...

The isolation of human oral mucosa/skin keratinocytes progenitor/stem cells is clinically important to regenerate epithelial tissues for the treatment of oral mucosa/skin defects. Researchers have attempted to isolate a keratinocyte progenitor/stem cell population usi ...

The isolation of hair follicle dermal papilla cells has become an important technique in the field of cutaneous stem cell biology. These cells can be used for a number of biological and translational purposes. They are studied to identify the cellular characteristics and molecular factors t ...

The increasing interest about stem cell research is linked to the promise of developing such treatments for many life-threatening, debilitating diseases and for cell replacement therapies. Among the various human tissues, skin represents a source characterized by great accessib ...

Recent studies revealed that mammalian dermis contains multipotent stem cells such as skin-derived precursors (SKPs). SKPs grow in suspension as spheres. In contrast, mesenchymal stem cells (MSCs) are adherent fibroblastic cells. Here, we describe the procedure to isolate MSCs under ...

Epidermal stem cells isolation struggle remains, mainly due to the yet essential requirement of well-defined approaches and markers. The herein proposed methodology integrates an assemblage of strategies to accomplish the enrichment of the interfollicular epidermal stem ce ...

Several different precursor populations participate in renewal and regeneration of the mammalian skin and hair follicle. Recently, we described the existence of multipotent dermal precursors that exhibit properties of stem cells, and reside in the mesenchymal compartment of the ...

Skin, as the body’s largest organ, has been extensively used to study adult stem cells. Most previous skin-related studies have focused on stem cells isolated from hair follicles and from keratinocytes. Here we present a protocol to isolate multipotent neural crest stem-like dermis-deriv ...

We developed protocols for isolation and characterization of mesenchymal progenitors from murine dermis. Our protocols are part of a more general isolation procedure starting with neonatal murine skin, which has been described in detail by U. Lichti and coauthors (Nat Protoc 3(5):799– ...

The discovery of human induced pluripotent stem cells (iPSCs) has provided a model system for studying early events during human development. Developmentally melanocytes originate from migratory neural crest cells that emerge from the neural plate during embryogenesis after a co ...

Melanoblasts are melanocyte precursors that are derived from neural crest cells (NCCs). Recently we showed that melanoblasts differentiate into not only pigmented melanocytes but also into other NCCs derivatives. Here, we describe methods for the isolation of melanoblasts from mo ...

This protocol describes an in vivo grafting approach to investigate the intrinsic long-term tissue reconstitutive capabilities of interfollicular keratinocyte stem cells and their committed progeny—the committed progenitors or transit amplifying and early different ...

The ability to grow keratinocyte stem cells (KSCs) in 3D culture is an important step forward for investigating the physiological properties of these cells. In the epidermis, KSCs are subject to various types of mechanical stress. To study the effects of mechanical stress on KSCs, monolayer cu ...

The epidermis of the skin and its appendages, such as the hair follicles, are formed and maintained by keratinocyte stem cells. Highly efficient and permanent genetic modifications are valuable tools to examine the multipotency and regenerative capacity of keratinocyte stem cells in s ...

Electrophoretic mobility shift assay and Western blot are simple, efficient, and rapid methods for the study of DNA–protein interactions and expression, respectively. Primary cultures and subcultures of epithelial cells are widely used for the production of tissue-engineered s ...

Gene expression analysis is a useful tool to study the molecular mechanisms underlying skin development and homeostasis. Here we describe a method that utilizes laser capture microdissection (LCM) to isolate RNAs from localized areas of skin, allowing the characterization of gene ex ...

Atomic force microscopy (AFM) can directly visualize single molecules in solution, which makes it an extremely powerful technique for carrying out studies of biological complexes and the processes in which they are involved. A recent development, called Recognition Imaging, allows ...

Expression of genes can be controlled by regulatory elements that are located at large genomic distances from their target genes (in cis), or even on different chromosomes (in trans). Regulatory elements can act at large genomic distances by engaging in direct physical interactions with th ...