Host Tissue Microenvironment Research Articles

Visualizing Staphylococcus aureus pathogenic membrane modification within the host infection environment by multimodal imaging mass spectrometry

Bacterial pathogens have evolved virulence factors to colonize, replicate, and disseminate within the vertebrate host. Although there is an expanding body of literature describing how bacterial pathogens regulate their virulence repertoire in response to environmental signals, it is challenging to directly visualize virulence response within the host tissue microenvironment. Multimodal imaging approaches enable visualization of host-pathogen molecular interactions. Here we demonstrate multimodal integration of high spatial resolution imaging mass spectrometry and microscopy to visualize Staphylococcus aureus envelope modifications within infected murine and human tissues. Data-driven image fusion of fluorescent bacterial reporters and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance imaging mass spectrometry uncovered S. aureus lysyl-phosphatidylglycerol lipids, localizing to select bacterial communities within infected tissue. Absence of lysyl-phosphatidylglycerols is associated with decreased pathogenicity during vertebrate colonization as these lipids provide protection against the innate immune system. The presence of distinct staphylococcal lysyl-phosphatidylglycerol distributions within murine and human infections suggests a heterogeneous, spatially oriented microbial response to host defenses.

Application of Mesenchymal Stem Cells in Inflammatory and Fibrotic Diseases.

Mesenchymal stem cells (MSCs) are multipotent stem cells that can be isolated from various tissues in the adult body. MSCs should be characterized by three criteria for regenerative medicine. MSCs must (1) adhere to plastic surfaces, (2) express specific surface antigens, and (3) differentiate into mesodermal lineages, including chondrocytes, osteoblasts, and adipocytes, in vitro. Interestingly, MSCs have immunomodulatory features and secrete trophic factors and immune receptors that regulate the microenvironment in host tissue. These specific and unique therapeutic properties make MSCs ideal as therapeutic agents in vivo. Specifically, pre-clinical and clinical investigators generated inflammatory and fibrotic diseases models, and then transplantation of MSCs into diseases models for therapeutic effects investigation. In this review, we characterize MSCs from various tissues and describe their applications for treating various inflammation and fibrotic diseases.

Platelet-Rich Plasma and Bone Marrow-Derived Mesenchymal Stromal Cells Prevent TGF-β1-Induced Myofibroblast Generation but Are Not Synergistic when Combined: Morphological in vitro Analysis

The persistence of activated myofibroblasts is a hallmark of fibrosis of many organs. Thus, the modulation of the generation/functionality of these cells may represent a strategical anti-fibrotic therapeutic option. Bone marrow-derived mesenchymal stromal cell (MSC)-based therapy has shown promising clues, but some criticisms still limit the clinical use of these cells, including the need to avoid xenogeneic compound contamination for ex vivo cell amplification and the identification of appropriate growth factors acting as a pre-conditioning agent and/or cell delivery vehicle during transplantation, thus enabling the improvement of cell survival in the host tissue microenvironment. Many studies have demonstrated the ability of platelet-rich plasma (PRP), a source of many biologically active molecules, to positively influence MSC proliferation, survival, and functionality, as well as its anti-fibrotic potential. Here we investigated the effects of PRP, murine and human bone marrow-derived MSCs, and of the combined treatment PRP/MSCs on in vitro differentiation of murine NIH/3T3 and human HDFα fibroblasts to myofibroblasts induced by transforming growth factor (TGF)-β1, a well-known pro-fibrotic agent. The myofibroblastic phenotype was evaluated morphologically (cell shape and actin cytoskeleton assembly) and immunocytochemically (vinculin-rich focal adhesion clustering, α-smooth muscle actin and type-1 collagen expression). We found that PRP and MSCs, both as single treatments and in combination, were able to prevent the TGF-β1-induced fibroblast-myofibroblast transition. Unexpectedly, the combination PRP/MSCs had no synergistic effects. In conclusion, within the limitations related to an in vitro experimentation, our study may contribute to providing an experimental background for supporting the anti-fibrotic potential of the combination PRP/MSCs which, once translated “from bench to bedside,” could potentially offer advantages over the single treatments.

Optimally Hierarchical Nanostructured Hydroxyapatite Coatings for Superior Prosthesis Biointegration.

Bone osteogenesis is a complex phenomenon dependent on numerous microenvironmental cues, with their synchrony regulating cellular functions, such as mechanical signaling, survival, proliferation, and differentiation, as well as controlled regional specification of skeletal progenitor cell fate. Therefore, obtaining a mechanistic understanding of cellular response to a microenvironment is now coming into intense focus, which will facilitate the design of programmable biomaterials for regenerative medicine. State-of-the-art nanomaterial synthesis and self-assembly processes yield complex structures that mimic surface properties, composition, and partially the morphology of the extracellular matrix. However, determining key structural properties that control cell attachment has been challenging and contradictory results are reported regarding the mechanisms and roll of nanostructured materials. Here, we significantly improve osteogenesis on bioinert substrates, demonstrating an exemplary organic-inorganic interface for superior prosthesis biointegration. We identify critical microscale hierarchical features that drastically enhance the cellular response to the same nanoscale architecture. It was observed that hierarchical morphologies, with a porosity above 80%, promote early-stage osteoinduction, as indicated by extensive coating ingrowth and nanofilopodia formation. We determined that cellular integration was mediated by two-way recognition of specific nano- and microtopographical cues between the host tissue and cellular microenvironment. This has allowed us to detail a set of determinant features for the nanofabrication of advanced prosthesis coatings that may ultimately improve implant longevity.

The Dynamic Identity of Intestinal Cancer Stem Cells

Elimination of self-renewing cancer stem cells (CSCs) is necessary to permanently eradicate malignant tissues. In a recent article in Nature, de Sousa e Melo etal. (2017) reveal that intestinal tumors can contain dynamic pools of functionally distinct CSC populations, which seem to interconvert depending on the host tissue microenvironment.

Mouse genomic screen reveals novel host regulator of metastasis

Tumor cells have to overcome challenges in the host tissue microenvironment to metastasize successfully to distant organs. In a recent Nature study, a genome-wide functional screen demonstrated that deficiency of the sphingosine-1-phoshate (S1P) transporter gene Spns2 in endothelium increased immune-mediated cell killing by T cells and natural killer (NK) cells, thereby suppressing metastatic colonization.

Mammary tissue microenvironment determines Tcell-dependent breast cancer-associated inflammation.

Although the importance of the host tissue microenvironment in cancer progression and metastasis has been established, the spatiotemporal process establishing a cancer metastasis-prone tissue microenvironment remains unknown. In this study, we aim to understand the immunological character of a metastasis-prone microenvironment in a murine 4T1 breast tumor model, by using the activation of nuclear factor-κb (NF-κB) in cancer cells as a sensor of inflammatory status and by monitoring its activity by bioluminescence imaging. By using a 4T1 breast cancer cell line stably expressing an NF-κB/Luc2 reporter gene (4T1 NF-κB cells), we observed significantly increased bioluminescence approximately 7 days after metastasis-prone orthotopic mammary fat-pad inoculation but not ectopic s.c. inoculation of 4T1 NF-κB cells. Such in vivo NF-κB activation within the fat-pad 4T1 tumor was diminished in immune-deficient SCID or nude mice, or T cell-depleted mice, suggesting the requirement of host T cell-mediated immune responses. Given the fat-pad 4T1 tumor expressed higher inflammatory mediators in a T cell-dependent mechanism compared to the s.c. tumor, our results imply the importance of the surrounding tissue microenvironment for inflaming tumors by collaborating with T cells to instigate metastatic spread of 4T1 breast cancer cells.

Metabolism Links Bacterial Biofilms and Colon Carcinogenesis

Bacterial biofilms in the colon alter the host tissue microenvironment. A role for biofilms in colon cancer metabolism has been suggested but to date has not been evaluated. Using metabolomics, we investigated the metabolic influence that microbial biofilms have on colon tissues and the related occurrence of cancer. Patient-matched colon cancers and histologically normal tissues, with or without biofilms, were examined. We show the upregulation of polyamine metabolites in tissues from cancer hosts with significant enhancement of N(1), N(12)-diacetylspermine in both biofilm-positive cancer and normal tissues. Antibiotic treatment, which cleared biofilms, decreased N(1), N(12)-diacetylspermine levels to those seen in biofilm-negative tissues, indicating that host cancer and bacterial biofilm structures contribute to the polyamine metabolite pool. These results show that colonic mucosal biofilms alter the cancer metabolome to produce a regulator of cellular proliferation and colon cancer growth potentially affecting cancer development and progression.

MP29-18 PREFABRICATION OF NEUROMUSCULAR JUNCTION FOR ACCELERATED RECOVERY OF MUSCLE FUNCTION

You have accessJournal of UrologyTrauma/Reconstruction: Ureter, Bladder, External Genitalia and Urotrauma II1 Apr 2015MP29-18 PREFABRICATION OF NEUROMUSCULAR JUNCTION FOR ACCELERATED RECOVERY OF MUSCLE FUNCTION In Kap Ko, Sang Jin Lee, John Jackson, Anthony Atala, and James Yoo In Kap KoIn Kap Ko More articles by this author , Sang Jin LeeSang Jin Lee More articles by this author , John JacksonJohn Jackson More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and James YooJames Yoo More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.621AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Surgical reconstruction involving muscle requires integration of nervous tissue for functional restoration. Failure of neural integration leads to atrophy and the tissue often becomes non-functional. Therefore, muscle innervation is a critical process in the recovery of function, this process is time sensitive and relies heavily on the host tissue microenvironment. Acceleration of nerve integration by facilitating the formation for neuromuscular junctions (NMJ) may be an option to restore muscle function after reconstruction. Toward this goal, we investigated whether muscle fibers cultured in vitro could form NMJ that would lead to rapid innervation to muscle tissue in vivo. METHODS Myotubes formed from muscle precursor cells (C2C12) were co-cultured with neuroblastoma cells (NG108), grown with conditioned medium derived from NG108, or treated with neurotrophic factors (agrin-containing medium). To confirm acetylcholine receptor (AChR) expression on the surface of myotubes, 10 μM of alpha-bungarotoxin (α-BTX) conjugated with fluorescent dye was added to the culture and visualized with a fluorescent microscope. AChR expressing myotubes were counted from randomly selected areas (n=9) in culture microplates. AChR expression on myotubes in a 3-D culture system was assessed. RESULTS Agrin treatment significantly increased the percentage of AChR expressing myotubes from 15% to 100%. Treatment with conditioned medium (CM) derived from NG108 cells in differentiation medium enhanced AChR expression by up to 50%; however, when CM was added to growth medium, AChR expression on the surface of myotubes was not affected. Treatment with agrin also increased AChR expression on myotubes grown in a 3-D culture system consisting of fibrin gel. CONCLUSIONS This study shows that expression of AchR can be controlled by the use of nerve, conditioned medium derived from neural cells or neurotrophic factors (agrin). These results suggest that formation of NMJ may allow for more effective neural integration and accelerate the recovery of muscle function. Furthermore, rapid integration of neural tissue would decrease the incidence of muscle atrophy and scarring. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e346 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information In Kap Ko More articles by this author Sang Jin Lee More articles by this author John Jackson More articles by this author Anthony Atala More articles by this author James Yoo More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Autotaxin and LPA1 and LPA5 receptors exert disparate functions in tumor cells versus the host tissue microenvironment in melanoma invasion and metastasis.

Autotaxin (ENPP2/ATX) and lysophosphatidic acid (LPA) receptors represent two key players in regulating cancer progression. The present study sought to understand the mechanistic role of LPA G protein-coupled receptors (GPCR), not only in the tumor cells but also in stromal cells of the tumor microenvironment. B16F10 melanoma cells predominantly express LPA5 and LPA2 receptors but lack LPA1. LPA dose dependently inhibited invasion of cells across a Matrigel layer. RNAi-mediated knockdown of LPA5 relieved the inhibitory effect of LPA on invasion without affecting basal invasion. This suggests that LPA5 exerts an anti-invasive action in melanoma cells in response to LPA. In addition, both siRNA-mediated knockdown and pharmacologic inhibition of LPA2 reduced the basal rate invasion. Unexpectedly, when probing the role of this GPCR in host tissues, it was found that the incidence of melanoma-derived lung metastasis was greatly reduced in LPA5 knockout (KO) mice compared with wild-type (WT) mice. LPA1-KO but not LPA2-KO mice also showed diminished melanoma-derived lung metastasis, suggesting that host LPA1 and LPA5 receptors play critical roles in the seeding of metastasis. The decrease in tumor cell residence in the lungs of LPA1-KO and LPA5-KO animals was apparent 24 hours after injection. However, KO of LPA1, LPA2, or LPA5 did not affect the subcutaneous growth of melanoma tumors. These findings suggest that tumor and stromal LPA receptors, in particular LPA1 and LPA5, play different roles in invasion and the seeding of metastasis.

Effects of different tissue microenvironments on gene expression in breast cancer cells.

In metastasis, circulating tumor cells penetrate the walls of blood vessels and enter the metastatic target tissue, thereby becoming exposed to novel and relatively unsupportive microenvironments. In the new microenvironments, the tumor cells often remain in a dormant state indefinitely and must adapt before they are able to successfully colonize the tissue. Very little is known about this adaptive process. We studied temporal changes in gene expression when breast cancer cells adapt to survive and grow on brain, bone marrow, and lung tissue maintained in an in vivo culture system, as models of the metastatic colonization of these tissues. We observed the transient activation of genes typically associated with homeostasis and stress during the initial stages of adaptation, followed by the activation of genes that mediate more advanced functions, such as elaboration of cell morphology and cell division, as the cells adapted to thrive in the host tissue microenvironment. We also observed the temporary induction of genes characteristic of the host tissue, which was particularly evident when tumor cells were grown on brain tissue. These early transient gene expression events suggest potential points of therapeutic intervention that are not evident in data from well-established tumors.

3D numerical simulation of avascular tumour growth: effect of hypoxic micro-environment in host tissue

A three-dimensional (3D) mathematical model of tumour growth at the avascular phase and vessel remodelling in host tissues is proposed with emphasis on the study of the interactions of tumour growth and hypoxic micro-environment in host tissues. The hybrid based model includes the continuum part, such as the distributions of oxygen and vascular endothelial growth factors (VEGFs), and the discrete part of tumour cells (TCs) and blood vessel networks. The simulation shows the dynamic process of avascular tumour growth from a few initial cells to an equilibrium state with varied vessel networks. After a phase of rapidly increasing numbers of the TCs, more and more host vessels collapse due to the stress caused by the growing tumour. In addition, the consumption of oxygen expands with the enlarged tumour region. The study also discusses the effects of certain factors on tumour growth, including the density and configuration of preexisting vessel networks and the blood oxygen content. The model enables us to examine the relationship between early tumour growth and hypoxic micro-environment in host tissues, which can be useful for further applications, such as tumour metastasis and the initialization of tumour angiogenesis.

Abstract 2871: Tumor-derived CCL2 promotes recruitment of CD11b/Gr1midmyeloid cells to sustain colorectal cancer liver metastasis.

Abstract Metastatic colorectal cancer (CRC) is one of the most prominent causes of cancer-related mortality. However, therapeutic options for patients with metastatic CRC are limited and development of new and more effective therapeutics aimed at downstaging the disease and prolonging survival is urgently required. Primary tumor dissemination to secondary sites, predominantly the liver, is a dynamic process dependent on the interactions between tumor cells and the host tissue microenvironment. Immune cells in the tumor microenvironment have been shown to play pivotal roles in the metastatic process but their pro-metastatic functions in CRC liver metastasis remains incompletely understood. To simulate liver metastasis, MC38 colon carcinoma cells were injected into the spleen of C57BL/6 mice. A population of CD11b/Gr1mid cells, distinct from other metastasis-associated myeloid cells previously described, increased dramatically during establishment of liver metastases. Adoptive transfer of bone marrow cells confirmed that these CD11b/Gr1mid cells were derived from bone marrow progenitor cells. MC38 tumor cells expressed high levels of CCL2 and serum levels were elevated in tumor-bearing mice compared to controls. Inhibition of tumor-derived CCL2 using shRNA, and absence of its cognate receptor in CCR2 KO mice significantly diminished CD11b/Gr1mid cell infiltration into tumor-bearing livers. Importantly, inhibition of CD11b/Gr1mid cell recruitment led to a marked reduction in tumor burden. Moreover, when CD11b/Gr1mid cells were depleted in a transgenic CD11b-DTR mouse model, a striking decrease in tumor vessel density was observed, with a concomitant reduction in tumor cell proliferation. Serum CCL2 concentrations were similarly elevated in CRC patients, including those who present metastasis, compared to normal controls. Furthermore, CD11b/CCR2 myeloid cells, analogous to the CD11b/Gr1mid population described in our mouse model, were identified in liver metastases of several CRC patients. Together, our studies demonstrate the importance of a distinct CD11b/Gr1mid myeloid subset in sustaining CRC liver metastasis and implicate a CCL2/CCR2-dependent pathway in their recruitment to metastatic sites. Our findings reinforce how tumor cells utilize host systems for successful propagation and identify a potential target for therapeutic manipulation. Citation Format: Su Yin Lim, Lei Zhao, Alex Gordon-Weeks, Thomas Tapmeier, Ruth Muschel. Tumor-derived CCL2 promotes recruitment of CD11b/Gr1mid myeloid cells to sustain colorectal cancer liver metastasis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2871. doi:10.1158/1538-7445.AM2013-2871

The effect of in vitro formation of acetylcholine receptor (AChR) clusters in engineered muscle fibers on subsequent innervation of constructs in vivo

Timely innervation of muscle tissue is critical in the recovery of function, and this time-sensitive process relies heavily on the host tissue microenvironment after implantation. However, restoration of muscle tissue mass and function has been a challenge. We investigated whether pre-forming acetylcholine receptor (AChR) clusters on engineered muscle fibers using an AChR cluster-inducing factor (agrin) prior to implantation would facilitate established contacts between implanted muscle tissues and nerves and result in rapid innervation of engineered muscle in vivo. We showed that agrin treatment significantly increased the formation of AChR clusters on culture differentiated myotubes (C2C12), enhanced contacts with nerves in vitro and in vivo, and increased angiogenesis. Pre-fabrication of AChR clusters on engineered skeletal muscle using a released neurotrophic factor can accelerate innervations following implantation in vivo. This technique has considerable potential for enhancing muscle tissue function.

Hypoxia-preconditioned mesenchymal stromal cells improve cardiac function in a swine model of chronic myocardial ischaemia

Cell loss during cardiac injection and hostility of the host-tissue microenvironment have the potential to diminish the overall effect of stem cell therapy. The purposes of this study were to evaluate the effect of a hypoxic preconditioning of mesenchymal stromal cells (MSC), to determine its safety and effectiveness, and to improve the efficacy of cell therapy using MSC in the setting of chronic myocardial ischaemia in swine. Myocardial ischaemia was induced by an ameroid constrictor. Human MSC were cultured under normoxic (20% O2) or hypoxic conditions (1.5% O2) before transplantation. One month after ischaemia, pigs were randomly assigned to saline injection (sham), and 1 × 10(6)/kg normoxic or hypoxic MSC transplantation into the ischaemic inferior-lateral zone. Twenty-seven pigs were operated on and the mortality rate was 33.3%. The remaining 18 animals were randomly assigned to sham (n = 4), normoxic (n = 8) or hypoxic MSC (n = 6) treatment. Global systolic (left ventricle ejection fraction, P = 0.04) and diastolic (E/Ea, P = 0.008) functions were increased in the hypoxic group compared with other groups. The peak of 2-dimensional longitudinal strain was less altered in the hypoxic group compared with other groups (P < 0.001). Haemodynamic data showed that dP/dT max was improved in the hypoxic group compared with the other group (P < 0.01). Capillary density was increased in the hypoxic group (P = 0.001). MSC density was significantly higher in the ischaemic zone in the hypoxic group (P < 0.01). MSC engraftment with hypoxic preconditioning significantly improves capillary density and cell survival, resulting in improvement in global, regional and diastolic left ventricular functions. This highlights the therapeutic potential of transplanting hypoxic-preconditioned MSC in the setting of chronic ischaemic heart failure.

Differentiating multipotent mesenchymal stromal cells generate factors that exert paracrine activities on exogenous MSCs: Implications for paracrine activities in bone regeneration

The mechanisms by which multipotent mesenchymal stromal cells (MSCs) contribute to tissue repair following transplantation into host tissues remains poorly understood. Current concepts suggest that, in addition to differentiation into cells of the host tissues, MSCs also generate trophic factors that modulate host tissue microenvironment to aid in the repair process. In this communication, we assessed whether factors secreted by MSCs undergoing osteogenic differentiation induce expression of osteoblast markers in exogenous MSCs as well as their migration. Murine MSCs were cultured in osteogenic medium, and at different time points, medium conditioned by the cells was collected and assessed for its effects on differentiation and migration of exogenous MSCs. In addition, we determined whether MSCs infused into mice femurs expressed genes encoding for factors predicted to play a role in paracrine activities. The results showed that MSCs maintained in osteogenic medium, secreted factors at specific time points that induced alkaline phosphatase activity (ALP) in exogenous MSCs as well as their migration. MSCs infused into mice femurs and retrieved at different days expressed genes that encoded predicted factors that play a role in cell differentiation and migration. Neutralizing antibodies to bone morphogenetic protein-2 (BMP-2) led to the decrease in ALP activity by exogenous MSCs. These data demonstrated that, as MSCs differentiate toward osteogenic lineage, they secrete factors that induce recruitment and differentiation of endogenous progenitors. These data reveal mechanisms by which donor MSCs may contribute to the bone reparative process and provide a platform for designing approaches for stem cell therapies of musculoskeletal disorders.

Using MKK4’s metastasis suppressor function to identify and dissect cancer cell–microenvironment interactions during metastatic colonization

Host tissue microenvironment plays key roles in cancer progression and colonization of secondary organs. One example is ovarian cancer, which colonizes the peritoneal cavity and especially the omentum. Our research indicates that the interaction of ovarian cancer cells with the omental microenvironment can activate a stress-kinase pathway involving the mitogen-activated protein kinase kinase 4 (MKK4). A combination of clinical correlative and functional data suggests that MKK4 activation suppresses growth of ovarian cancer cells lodged in omentum. These findings prompted us to turn our focus to the cellular composition of the omental microenvironment and its role in regulating cancer growth. In this review, in addition to providing an overview of MKK4 function, we highlight a use for metastasis suppressors as a molecular tool to study cancer cell interaction with its microenvironment. We review features of the omentum that makes it a favorable microenvironment for metastatic colonization. In conclusion, a broader, evolutionary biology perspective is presented which we believe needs to be considered when studying the evolution of cancer cells within a defined microenvironment. Taken together, this approach can direct new multi-dimensional lines of research aimed at a mechanistic understanding of host tissue microenvironment, which could be used to realize novel targets for future research.

Use of Porous Polyethylene Implants in Nasal Reconstruction

ABSTRACT Introduction Patients undergoing complex rhinoplasty, following previous trauma or cartilage deteriorating disease processes, present a surgical challenge. The availability of septal cartilage maybe limited, and alternative graft materials should be considered. Methods A retrospective study of our experience using the porous polyethylene (Medpor®) implant in nasal reconstruction was conducted. The indications for surgery are reviewed and the surgical outcomes noted. We also review the literature on available nasal implants and discuss their advantages and disadvantages. Results A total of 22 Medpor® nasal implants were used for augmentation rhinoplasty over a 6 and half years period. Three patients experienced a surgical complication, including one slipped dorsal implant, and two patients with graft extrusion. Both of these two patients had concomitant disease processes that would have resulted in a poor tissue microenvironment, which may have contributed to their graft failures. Conclusion In our experience, the Medpor® implant has proved very successful in patients requiring complex rhinoplasty, in a normal host tissue microenvironment.

Surgical Transplantation of Mouse Neural Stem Cells into the Spinal Cords of Mice Infected with Neurotropic Mouse Hepatitis Virus

Mice infected with the neurotropic JHM strain of mouse hepatitis virus (MHV) develop pathological and clinical outcomes similar to patients with the demyelinating disease Multiple Sclerosis (MS). We have shown that transplantation of NSCs into the spinal cords of sick mice results in a significant improvement in both remyelination and in clinical outcome. Cell replacement therapies for the treatment of chronic neurologic diseases are now a reality and in vivo models are vital in understanding the interactions between the engrafted cells and host tissue microenvironment. This presentation provides an adapted method for transplanting cells into the spinal cord of JHMV-infected mice. In brief, we provide a procedure for i) preparation of NSCs prior to transplant, ii) pre-operative care of mice, iii) exposure of the spinal cord via laminectomy, iv) stereotactic injection of NSCs, and iv) post-operative care.

Surgical Transplantation of Mouse Neural Stem Cells into the Spinal Cords of Mice Infected with Neurotropic Mouse Hepatitis Virus

Mice infected with the neurotropic JHM strain of mouse hepatitis virus (MHV) develop pathological and clinical outcomes similar to patients with the demyelinating disease Multiple Sclerosis (MS). We have shown that transplantation of NSCs into the spinal cords of sick mice results in a significant improvement in both remyelination and in clinical outcome. Cell replacement therapies for the treatment of chronic neurologic diseases are now a reality and in vivo models are vital in understanding the interactions between the engrafted cells and host tissue microenvironment. This presentation provides an adapted method for transplanting cells into the spinal cord of JHMV-infected mice. In brief, we provide a procedure for i) preparation of NSCs prior to transplant, ii) pre-operative care of mice, iii) exposure of the spinal cord via laminectomy, iv) stereotactic injection of NSCs, and iv) post-operative care.