Human Hair Follicles: “Bulging” with Neural Crest–Like Stem Cells

Several studies have reported the existence of precursor cells residing within various adult tissues that appear to either retain or recapitulate features of neural crest stem cells (NCSCs). In rodents, unique populations of both epidermal and dermal cells, resident within hair follicles, exhibit such characteristics, although the existence of equivalent NCSC-like cells in human tissues has remained uncertain. In this issue, Yu et al. show that NCSC-like cells also reside within the bulge region of adult human hair follicles

Presence of a loner strain maintains cooperation and diversity in well-mixed bacterial communities

Cooperation and diversity abound in nature despite cooperators risking exploitation from defectors and superior competitors displacing weaker ones. Understanding the persistence of cooperation and diversity is therefore a major problem for evolutionary ecology, especially in the context of well-mixed populations, where the potential for exploitation and displacement is greatest. Here we demonstrate that a “loner effect”, described by economic game theorists, can maintain cooperation and diversity in real-world biological settings. We use mathematical models of public-good-producing bacteria to show that the presence of a loner strain, which produces an independent but relatively inefficient good, can lead to rock-paper-scissor dynamics, whereby cooperators outcompete loners, defectors outcompete cooperators, and loners outcompete defectors. These model predictions are supported by our observations of evolutionary dynamics in well-mixed experimental communities of the bacterium Pseudomonas aeruginosa. We find that the coexistence of cooperators and defectors, which respectively produce and exploit the iron-scavenging siderophore pyoverdine, is stabilized by the presence of loners with an independent iron-uptake mechanism. Our results establish the loner effect as a simple and general driver of cooperation and diversity in environments that 40 would otherwise favour defection and the erosion of diversity.Publisher PDFPeer reviewe

Defining The Role of Primary Cilia on Skin Derived Precursors.

Isolated in 2001, Skin Derived Precursors (SKPs) represent a novel population of multipotent stem cells1,2 residing at the base of hair follicle where they play a key role in defining the physiology and regeneration capacity of hair follicles and skin (Biernaskie et al. 2009). In order to understand how SKPs behavior is regulated in the skin, we asked whether primary cilia, microtubule bundles projected from the cell surface that transmit chemical signals between cells4, are present in the dermal papilla and dermal sheath in-vivo and in the population of isolated SKPs. In our investigation, we performed immunohistochemistry for acetylated tubulin and showed that SKPs do have cilia. We then tested whether these cilia can be elongated following treatments of lithium chloride on these dermal precursors. We then asked whether elongation of cilia has an effect on the self-renewal capacity of SKPs and whether drugs elongating cilia can work synergistically with Platelet Derived Growth Factor (PDGF), a growth factor that we have previously shown to improve cell growth and cell division. Experiments assessing the self-renewal capacity of SKPs suggested significant  increase in the diameter and the number of spherical presence when treated with lithium chloride and when lithium chloride is added in combination with PDGFβ when compared to PDGFβ alone. This suggests that signaling in cilia may influence PDGF signals causing an enhanced effect on SKP proliferation. Further experiments including knocking down primary cilia by blocking the transcription of ciliary protein using shRNA and in-vivo transplantations of lithium chloride and PDGF treated SKPs in a hair follicle formation assays will be executed to understand the key roles of primary cilia on SKPs. These studies will ultimately aim to answer whether drugs affecting cilia can function as potential therapeutic targets for autologous adult stem cell based therapies

METABOLISM OF SKIN DERIVED PRECURSOR (SKP) CELLS IN STIRRED SUSPENSION BIOREACTORS

INTRODUCTION More than six million people suffer from burn injuries every year. These injuries can result in psychological trauma, disabilities and permanent disfigurement. A common treatment for burn victims is an autologous graft surgery in which skin is transplanted from a healthy part of the body to the injury site (i.e. split thickness skin graft). This graft, however, does not contain functional dermal tissue, hair follicles or glands, often causing graft contraction, chronic irritation, and unnatural in appearance. We have hypothesized that skin-derived precursor (SKP) cells, a multipotent dermal stem cell that resides within skin hair follicles, can be utilized in conjunction with split thickness skin grafts to improve their function and minimize irritation. SKPs have a high proliferative potential and need to be expanded in a well-controlled, standardized culture environment before they can be utilized in clinical treatments [1]. It is essential to optimize the expansion of SKP cells in order to generate a bioprocess capable of producing enough cells for a clinical setting. METHODS -qNUTR = ∆[NUTR]/Int(Xv)dtThe specific uptake and production rates were calculated for cells cultured in static T-Flask environments and stirred suspension bioreactors run at 40, 60, 80, and 100 rpm. Equation 1 was used to calculate specific rates (qNUTR). The integral change in viable cells (Int(Xv)dt) was calculated using a numerical trapezoid approximation, and the change in nutrient concentration (∆[NUTR]) was measured using the Nova Bioprofile 100+ analyzer. SKP cells were taken from a 68 year old female. RESULTS The specific uptake rates of glucose and glutamine and specific production rates of lactate and ammonia have been determined under altered cell culture environments for SKP cells (Figure1). This provides details into nutrient limitations and cell metabolic behaviours needed to access parameters to guide our bioprocess design and development of robust expansion protocols. DISCUSSION AND CONCLUSIONS Low levels of oxygen and nutrients result in significant changes to cell growth rates [2].  We were able to conclude that SKP cell growth is not limited by the glucose or glutamine concentrations in the media, and lactate and ammonia do not reach toxic levels. It is interesting to note that the stirred suspension environment does appear to have an effect on the specific consumption rates of glucose and glutamine. According to these initial results, we are predicting that under shear stress environments, SKP cells are changing their metabolic behaviour to allow more glucose to convert to pyruvate and enter the TCA cycle. The specific production rates of lactate and ammonia, however, do not follow the same patterns. Further validation and reasons behind these differences need to be investigated

Development of an alternative harvesting method using pH to detach adherent cells from microcarriers

Peripheral nerve injuries are common in Canada, affecting 2.8% of trauma patients treated every year. Current repair strategies are inadequate and repair is often suboptimal with only 25% of patients recovering full motor function and only 3% regaining full sensory function. Because of this, the field is turning toward regenerative medicine to develop a cellular therapy using Schwann cells to repair injured nerves. Schwann cells differentiated from skin derived precursors (SKP-SCs) are a promising cell type as they are easily obtained and allow for autologous therapy. To be able to generate clinically relevant numbers of SKP-SCs, bioreactors need to be used. Since SKP-SCs are an adherent cell type, to be expanded in suspension bioreactors, small spherical beads known as microcarriers need to be used. Our lab has previously shown that these SKP-SCs readily attach to the microcarriers and grow in stirred suspension bioreactors. We have also shown that by controlling the culture parameters, we can increase the maximum cell density compared to conventional static culture methods. One of the biggest hurdles that remains is an efficient harvesting method that can be scaled up to clinical applications. Current cell detachment protocols use enzymatic based solutions to remove the cells from the surface of the microcarriers. These methods work well in removing the cells, however, they are very labour intensive as they require many washing steps and taking the reactors offline. Therefore, we looked into an alternative method for the detachment of SKP-SCs from microcarriers that will allow for an inline detachment process. This new method is based on previous research done in our lab using high pH solutions to dissociate aggregates. First we investigated the detachment efficiency in static. Cells were cultured in 6-well plates until confluency and then harvested with solutions ranging from pH 8-9.5. With a pH of 9 and an incubation time of 30 minutes, we were able to recover 75% of cells when compared to traditional enzymatic harvesting. Following this we performed a qualitative analysis on the detachment of the SKP-SCs from the microcarriers to determine if this method has potential. Small 3mL samples were taken and solutions with pHs 8.5, 9, and 9.5 were added and incubated for 30 minutes and agitated every 5 minutes. We found that the cells detached with a high efficiency after 30 minutes with a pH of only 8.5. This was then quantified while maintaining a viability of above 90%. Following this we tested this method in harvesting full 125mL bioreactors. We evaluated different pH, agitation rates, and incubation times. We also assessed the ability of the cells to reattach to microcarriers and continue to expand over several serial passages to ensure there were no negative effects on the cells. Lastly we looked at using this method in our controlled bioreactors to increase the pH without the addition of anything else. Based on our results, increasing the pH of the culture medium can detach the SKP-SCs from microcarriers at a pH as low as 8.5 which allows for minimal cell damage while still detaching cells. We also noted that when the pH gets too high (\u3e9.5), the microcarriers begin to clump together causing large aggregates of microcarriers which could lead to clogging during the filtration steps. With increasing agitation, higher recovery efficiencies can be achieved indicating that this method of cell detachment has potential for large volume processes

Factors Within the Endoneurial Microenvironment Act to Suppress Tumorigenesis of MPNST

Background: Deciphering avenues to adequately control malignancies in the peripheral nerve will reduce the need for current, largely-ineffective, standards of care which includes the use of invasive, nerve-damaging, resection surgery. By avoiding the need for en bloc resection surgery, the likelihood of retained function or efficient nerve regeneration following the control of tumor growth is greater, which has several implications for long-term health and well-being of cancer survivors. Nerve tumors can arise as malignant peripheral nerve sheath tumors (MPNST) that result in a highly-aggressive form of soft tissue sarcoma. Although the precise cause of MPNST remains unknown, studies suggest that dysregulation of Schwann cells, mediated by the microenvironment, plays a key role in tumor progression. This study aimed to further characterize the role of local microenvironment on tumor progression, with an emphasis on identifying factors within tumor suppressive environments that have potential for therapeutic application.Methods: We created GFP-tagged adult induced tumorigenic Schwann cell lines (iSCs) and transplanted them into various in vivo microenvironments. We used immunohistochemistry to document the response of iSCs and performed proteomics analysis to identify local factors that might modulate divergent iSC behaviors.Results: Following transplant into the skin, spinal cord or epineurial compartment of the nerve, iSCs formed tumors closely resembling MPNST. In contrast, transplantation into the endoneurial compartment of the nerve significantly suppressed iSC proliferation. Proteomics analysis revealed a battery of factors enriched within the endoneurial compartment, of which one growth factor of interest, ciliary neurotrophic factor (CNTF) was capable of preventing iSCs proliferation in vitro.Conclusions: This dataset describes a novel approach for identifying biologically relevant therapeutic targets, such as CNTF, and highlights the complex relationship that tumor cells have with their local microenvironment. This study has significant implications for the development of future therapeutic strategies to fight MPNSTs, and, consequently, improve peripheral nerve regeneration and nerve function

Investment and Allocation of nectar Production in an Animal-Pollinated Plant

A plant\u27s ability to attract animal pollinators should be influenced by two important factors: the amount of available resources that is invested to the production of rewards such as nectar, and the individual\u27s size or resource state. Using Chamerion (= Epilobium) angustifolium (Onagraceae) as a model, I investigate: (1) selection pressures that determine the evolutionarily stable investment to nectar production, and (2) allocation tactics that vary with an individual\u27s current resource state. A conceptual model is presented, in which a plant\u27s attractiveness to pollinators increases with relative investment to nectar production (\u27social competition\u27). The evolutionarily stable investment strategy occurs where the expense of further nectar production outweighs the potential disadvantage of being slightly less attractive than competitors. The model may explain why C. angustifolium inflorescences produce, on average, such large amounts of nectar. Within populations, however, individuals varied greatly in the amount of nectar offered to pollinators, mainly due to variation in floral display size (number of open flowers-a correlate of resource state). Given that large, attractive inflorescences maximize pollen export by limiting the amount of pollen removed by each visitor, I predicted a size-dependent distribution of nectar within the vertical inflorescences of C. angustifolium. As predicted, small inflorescences distributed nectar nearly evenly among (lower) female- and (upper) male-phase flowers, whereas larger inflorescences allocated extra nectar to female-phase flowers. In experimental inflorescences, I distributed the same volume of nectar to mimic the \u27large\u27 and \u27small\u27 allocation types. Nectar-foraging bumblebees visited a mean of 3.2 fewer male-phase flowers on the \u27large\u27, relative to \u27small\u27 type, as expected if the nectar distribution of large displays functions to limit pollen removal during individual visits. I propose that the nectar gradient may adaptively mediate the schedule of pollen removal by manipulating the patch departure behaviour of pollinators

Biocomposite nanofiber matrices to support ECM remodeling by human dermal progenitors and enhanced wound closure

Cell-based therapies have recently been the focus of much research to enhance skin wound healing. An important challenge will be to develop vehicles for cell delivery that promote survival and uniform distribution of cells across the wound bed. These systems should be stiff enough to facilitate handling, whilst soft enough to limit damage to newly synthesized wound tissue and minimize patient discomfort. Herein, we developed several novel modifiable nanofibre scaffolds comprised of Poly (ε-caprolactone) (PCL) and gelatin (GE). We asked whether they could be used as a functional receptacle for adult human Skin-derived Precursor Cells (hSKPs) and how naked scaffolds impact endogenous skin wound healing. PCL and GE were electrospun in a single facile solvent to create composite scaffolds and displayed unique morphological and mechanical properties. After seeding with adult hSKPs, deposition of extracellular matrix proteins and sulphated glycosaminoglycans was found to be enhanced in composite grafts. Moreover, composite scaffolds exhibited significantly higher cell proliferation, greater cell spreading and integration within the nanofiber mats. Transplantation of acellular scaffolds into wounds revealed scaffolds exhibited improvement in dermal-epidermal thickness, axonal density and collagen deposition. These results demonstrate that PCL-based nanofiber scaffolds show promise as a cell delivery system for wound healing

Enhanced Expansion and Sustained Inductive Function of Skin-Derived Precursor Cells in Computer-Controlled Stirred Suspension Bioreactors

Endogenous dermal stem cells (DSCs) reside in the adult hair follicle mesenchyme and can be isolated and grown in vitro as self-renewing colonies called skin-derived precursors (SKPs). Following transplantation into skin, SKPs can generate new dermis and reconstitute the dermal papilla and connective tissue sheath, suggesting they could have important therapeutic value for the treatment of skin disease (alopecia) or injury. Controlled cell culture processes must be developed to efficiently and safely generate sufficient stem cell numbers for clinical use. Compared with static culture, stirred-suspension bioreactors generated fivefold greater expansion of viable SKPs. SKPs from each condition were able to repopulate the dermal stem cell niche within established hair follicles. Both conditions were also capable of inducing de novo hair follicle formation and exhibited bipotency, reconstituting the dermal papilla and connective tissue sheath, although the efficiency was significantly reduced in bioreactor-expanded SKPs compared with static conditions. We conclude that automated bioreactor processing could be used to efficiently generate large numbers of autologous DSCs while maintaining their inherent regenerative function

A common rule for decision-making in animal collectives across species

A diversity of decision-making systems has been observed in animal collectives. In some species, choices depend on the differences of the numbers of animals that have chosen each of the available options, while in other species on the relative differences (a behavior known as Weber's law) or follow more complex rules. We here show that this diversity of decision systems corresponds to a single rule of decision-making in collectives. We first obtained a decision rule based on Bayesian estimation that uses the information provided by the behaviors of the other individuals to improve the estimation of the structure of the world. We then tested this rule in decision experiments using zebrafish (Danio rerio), and in existing rich datasets of argentine ants (Linepithema humile) and sticklebacks (Gasterosteus aculeatus), showing that a unified model across species can quantitatively explain the diversity of decision systems. Further, these results show that the different counting systems used by animals, including humans, can emerge from the common principle of using social information to make good decisions