Hair follicle dermal cells repopulate the mouse haematopoietic system

Skin and hair follicle stem cell biology is the focus of increasing interest, not least because the adult hair follicle has well defined dermal and epithelial populations that display distinct developmental properties. Recent evidence suggests that a number of adult cell populations have much broader stem cell capabilities than previously thought. To examine whether this applied to the hair follicle, and with a view to developing the follicle as a stem cell model system we investigated whether adult hair follicles were capable of demonstrating haematopoietic stem cell activity. To investigate haematopoietic activity in hair follicles we first used in vitro haematopoietic colony assays. This demonstrated that rodent hair follicle end bulbs as well as micro-dissected dermal papilla and dermal sheath cells actively produced cells of erythroid and myeloid lineages but that follicle epithelial cells did not. As a more stringent test, we then transplanted cultured dermal papilla or dermal sheath cells from transgenically marked donor mice into lethally irradiated recipient mice and observed multi-lineage haematopoietic reconstitution when assayed at intervals of up to one year. Colony assays from bone marrow of primary recipients revealed that over 70% of clonogenic precursors were derived from donor hair follicle cells. When bone marrow from primary mice was harvested and used to repopulate secondary myeloablated recipients, multi-lineage haematopoietic engraftment was observed. Our data show that dermal but not epidermal compartments of the adult hair follicle have much broader stem cell activities than previously described. Although the treatment for many forms of blood disorder, such as leukemia, often requires transplantation of haematopoietic stem cells (HSC), their availability can be rate limiting. Given its easy accessibility, our identification of the hair follicle as a source of extramedullary haematopoietic stem cell activity makes it an attractive potential source for blood stem cell therapeutics and highlights its value as a model system in adult stem cell biology

Cultured human and rat tooth papilla cells induce hair follicle regeneration and fiber growth

The mesenchymal–epithelial interactions that characterize the early stages of tooth and hair follicle morphogenesis share certain similarities, and there is increasing evidence that mesenchymal cells derived from both mature structures retain interactive and stem cell-like properties. This study aimed to gauge the cross-appendage inductive capabilities of cultured tooth dental papilla (or pulp) cells from different species and ages of donor. Adult human and juvenile rat tooth papilla cells were implanted into surgically inactivated hair follicles within two different microenvironments. The human cells interacted with follicle epithelium to regenerate new end bulbs and create multiple differentiated hair fibers. Rodent tooth dental cells also induced new epithelial matrix structures and stimulated de novo hair formation. However, in many instances they also elicited mineralization and bone formation, a phenomenon that appeared to relate to their donor's age; the type of tooth of origin; and the host environment. Taken together, this study reveals that cultured dental papilla cells from postnatal mammals (adult, juvenile, and newborn) retain inductive molecular signals that must be common to both hair and teeth follicles. It highlights the stem cell-like qualities and morphogenetic abilities of tooth and hair follicle cells from mature humans, and their capacity for cross-appendage and interspecies communication and interaction. Besides the developmental implications, the present findings have relevance for stem cell biology, hair growth, tissue repair, and other biotechnologies. Moreover, the critical importance of considering the local microenvironment in which different cells/tissues are naturally or experimentally engineered is firmly demonstrated

Tissue engineering matrixes by emulsion templating

Foams containing poly(lactic acid) (PLA) have been prepared from the corresponding macromonomers using high internal phase emulsions (HIPEs) as templates to create the porous structure. The resulting PolyHIPE foams have been investigated by scanning electron microscopy and the influence of diluent type, either reactive (styrene or methyl methacrylate (MMA)) or unreactive (toluene), on foam morphology has been determined. The morphology was found to depend on the type of diluent: styrene resulted in cellular materials at low PLA levels but tended to give structures characteristic of phase separation at higher PLA content; MMA yielded porous cellular materials at all levels of PLA; toluene tended to give soft materials that collapsed on drying. The swelling of the foam materials in toluene, isopropyl alcohol (IPA) and water was also investigated. It was found that most of the foams swelled to a relatively low extent in all solvents, apart from that of low PLA content made with toluene. This was ascribed to a low crosslinking level and high porosity of this material. Cell and tissue growth studies on the resulting foams, as well as analogues prepared from poly(epsilon-caprolactone) (PCL) macromonomers, using whole chicken embryo explants, rat skin explants or individual human skin cells, indicated excellent biocompatibility of all foams over the length of each experiment (maximum length 6 days). Comparative studies indicated that cells adhered to PCL-based materials more rapidly than their PLA-containing counterparts

In vivo induction of hair growth by dermal cells isolated from hair follicles after extended organ culture

Successful hair follicle organ culture has been established for some time, but hair growth in vitro is limited and generally terminates prematurely in comparison with in vivo. The reasons why growth stops in culture are as yet unknown. In this investigation, adult rat vibrissa follicles for which growth in culture is limited to about 10 d, were maintained in vitro for a minimum of 20 d after the hair shaft stopped growing. The pattern of fiber growth and long-term follicle pathology reflected the initial hair cycle stage at the time of isolation. Furthermore, there was evidence that a group of follicles put into culture when in late anagen were attempting to cycle in vitro. Microscopy showed that, in spite of widespread pathologic changes to the follicle epithelium, dermal cells in the follicle showed remarkable resilience. Their viability was confirmed when primary cell cultures were established from isolated dermal tissue. These cells labeled positively for -smooth muscle actin, an established marker of hair follicle dermal cell phenotype in vitro. Moreover, isolated dermal tissue induced hair growth when implanted into inactivated hair follicles in vivo. These data confirm that the cessation in hair growth is not due to a loss of the inductive capacity in the dermal component. Long-term organ culture may provide opportunities to investigate factors that are expressed or lost during hair growth cessation. In addition it may be possible to develop this method further to obtain a reliable and predictable model of hair follicle cycling in vitro

Development of the mouse dermal adipose layer occurs independently of subcutaneous adipose tissue and is marked by restricted early expression of FABP4

The laboratory mouse is a key animal model for studies of adipose biology, metabolism and disease, yet the developmental changes that occur in tissues and cells that become the adipose layer in mouse skin have received little attention. Moreover, the terminology around this adipose body is often confusing, as frequently no distinction is made between adipose tissue within the skin, and so called subcutaneous fat. Here adipocyte development in mouse dorsal skin was investigated from before birth to the end of the first hair follicle growth cycle. Using Oil Red O staining, immunohistochemistry, quantitative RT-PCR and TUNEL staining we confirmed previous observations of a close spatio-temporal link between hair follicle development and the process of adipogenesis. However, unlike previous studies, we observed that the skin adipose layer was created from cells within the lower dermis. By day 16 of embryonic development (e16) the lower dermis was demarcated from the upper dermal layer, and commitment to adipogenesis in the lower dermis was signalled by expression of FABP4, a marker of adipocyte differentiation. In mature mice the skin adipose layer is separated from underlying subcutaneous adipose tissue by the panniculus carnosus. We observed that the skin adipose tissue did not combine or intermix with subcutaneous adipose tissue at any developmental time point. By transplanting skin isolated from e14.5 mice (prior to the start of adipogenesis), under the kidney capsule of adult mice, we showed that skin adipose tissue develops independently and without influence from subcutaneous depots. This study has reinforced the developmental link between hair follicles and skin adipocyte biology. We argue that because skin adipocytes develop from cells within the dermis and independently from subcutaneous adipose tissue, that it is accurately termed dermal adipose tissue and that, in laboratory mice at least, it represents a separate adipose depot

Microenvironmental reprogramming by three-dimensional culture enables dermal papilla cells to induce de novo human hair-follicle growth.

De novo organ regeneration has been observed in several lower organisms, as well as rodents; however, demonstrating these regenerative properties in human cells and tissues has been challenging. In the hair follicle, rodent hair follicle-derived dermal cells can interact with local epithelia and induce de novo hair follicles in a variety of hairless recipient skin sites. However, multiple attempts to recapitulate this process in humans using human dermal papilla cells in human skin have failed, suggesting that human dermal papilla cells lose key inductive properties upon culture. Here, we performed global gene expression analysis of human dermal papilla cells in culture and discovered very rapid and profound molecular signature changes linking their transition from a 3D to a 2D environment with early loss of their hair-inducing capacity. We demonstrate that the intact dermal papilla transcriptional signature can be partially restored by growth of papilla cells in 3D spheroid cultures. This signature change translates to a partial restoration of inductive capability, and we show that human dermal papilla cells, when grown as spheroids, are capable of inducing de novo hair follicles in human skin