细胞功能测定

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 ...

Before leaving the site of transcription, newborn messenger RNAs (mRNAs) become associated with a number of different proteins. How these large messenger ribonucleoprotein (mRNP) complexes then move through the dense nucleoplasm to reach the nuclear periphery has been a fascinati ...

Molecular combing is a process whereby single DNA molecules bind by their extremities to a silanised surface and are then uniformly stretched and aligned by a receding air/water interface (1). This method, with a high resolution ranging from a few kilobases to megabases, has many applications ...

The Miller chromatin spreading technique for electron microscopic visualization of gently dispersed interphase chromatin has proven extremely valuable for analysis of genetic activities in vivo. It provides a unique view of transcription and RNA processing at the level of indiv ...

Gene expression in eukaryotic cells is a multi-step process. Many of the steps are both co-ordinated and quality controlled. For example, transcription is closely coupled to pre-messenger RNA (mRNA)–protein assembly, pre-mRNA processing, surveillance of the correct synthesis of me ...

The mechanical properties of the interphase nucleus have important implications for cellular function and can reflect changes in nuclear envelope structure and/or chromatin organization. Mutations in the nuclear envelope proteins lamin A and C cause several human diseases, such ...

Understanding the physical properties of the cell nucleus is critical for developing a deeper understanding of nuclear structure and organization as well as how mechanical forces induce changes in gene expression. We use micropipette aspiration to induce large, local deformations ...