Special Issue:
Challenges and Opportunities in Tissue Engineering
Christian Elmshaeuser
Stem Cell Therapy Division, Institute of Medical Microbiology, University of Giessen, Giessen Germany & International Senior Professional Institute (ISPI) e.V. Giessen, Germany. 2Biochemistry Institute, University of Giessen, Giessen, Germany.
Ina Zoeller
Stem Cell Therapy Division, Institute of Medical Microbiology, University of Giessen, Giessen Germany & International Senior Professional Institute (ISPI) e.V. Giessen, Germany. 2Biochemistry Institute, University of Giessen, Giessen, Germany.
Darisuren Anhlan
Biochemistry Institute, University of Giessen, Giessen, Germany.
Ewald Beck
Biochemistry Institute, University of Giessen, Giessen, Germany.
Bruno Peault
INSERM U506, Groupe Hospitalier Paul Brousse, Villejuif Cedex, France & Orthopaedic Hospital Research Center and Broad Stem Cell Research Center, David Geffen School of Medicine, University of California, Los Angeles, LA, United States & MRC Centre Institute for Regenerative Medicine and Centre for Cardiovascular Science, University of Edinburgh, Edinburgh, United Kingdom.
Olivier Tabary
Sorbonne Université, INSERM UMR S 938, Centre de Recherche Saint Antoine (CRSA), Paris, France.
Una Chen
Stem Cell Therapy Division, Institute of Medical Microbiology, University of Giessen, Giessen Germany & International Senior Professional Institute (ISPI) e.V. Giessen, Germany. ; 5Research Experience of Stem Cells in Europe Society (RESCUES) e.V., Giessen, Germany.
Abstract
Background-Purpose of this study: Tissue-specific stem cell lines are useful tools for cell biology studies. Information on respiratory tissue cell lines is limited. A doxycycline-regulated epithelial precursor cell line was established from the lung tissue of a tTAxSV40 Tag double transgenic mouse. In this study, we have characterized this cell line in vitro & in vivo, and found to mimic a rare subpopulation of club- and pneumocyte type II-dual cells.
Methods: It was partially characterized using cell viability and death assays, H3-thymidine incorporation assay, chloride efflux assay, Western blotting of proteins secreted, RT-PCR assays for RNA isolated. In addition, immune–deficient SCID mice were used as hosts for implantation of this precursor cell line, and feed with/without doxycycline containing water. Immunofluorescent typing using different antibodies were used to characterize the implanted lung.
Results: This cell line was found to mimic a rare subpopulation of club- and pneumocyte type II- dual cells with multiple phenotypes. Cell growth was doxycycline-regulated and observed only when doxycycline was omitted from the medium or present at concentrations up to 1 µg/ml, higher concentrations were inhibitory. ACT+ ciliated cells were found upon implantation into immune-deficient mice, in addition. Cell growth was doxycycline-regulated in vitro. When transplanted subcutaneously into immune–deficient mice, these cells migrated to the lung to form organized chimeric structures of donor and host origins, with club cells in the terminal bronchioles, ACT+ ciliated cells along the epithelial lining, and pneumocyte type II-cells in the alveolar interstices. No such homing of donor cells to the lung was observed when the implanted mice were fed doxycycline-containing water.
Discussions-Conclusions: This lung stem cell line might be able to provide us with an insight into the differentiation pathway of lung epithelial cells as well as with some understanding of the nature of air trophic-pulmonary epithelial cells. The results of this study underline the possibility of a future application for somatic (stem / precursor) cells in tissue replacement and tissue engineering of the damaged lung. Its ability to secrete and deliver soluble protein, might be a potential novel way for drug delivery. In addition, stem cells are thought to proliferate and differentiate in response to a deficiency or as a result of injury. Successful migration to the target organ and subsequent maturation of these precursors could be attributed to a requirement of lung stem cells to search for an aerated environment. Our findings challenge some current concepts of stem cell biology.This lung stem cell line may become a rich source of cells for tissue engineering and cell-based therapy for lung injury. The route and protocol established for cell introduction into the lung may provide a novel alternative to delivery of soluble protein substances through the airways. This lung stem cell line might also be modified to provide models for screening drugs against respiratory infection.
Alan B. Moy
Cellular Engineering Technologies, Inc. Coralville, IA, 52241; John Paul II Medical Research Institute, Coralville, IA 52241
Anant Kamath
Cellular Engineering Technologies, Inc. Coralville, IA, 52241
Sara Ternes
Cellular Engineering Technologies, Inc. Coralville, IA, 52241
Jay Kamath
John Paul II Medical Research Institute, Coralville, IA 52241
Abstract
Induced pluripotent stem cells (iPSC) represent a potentially exciting regenerative-medicine cell therapy for several chronic conditions such as macular degeneration, soft tissue and orthopedic conditions, cardiopulmonary disease, cancer, neurodegenerative disorders and metabolic disorders. The field of iPSC therapeutics currently exists at an early stage of development. There are several important stakeholders that include academia, industry, regulatory agencies, financial institutions and patients who are committed to advance the field. Yet, unlike more established therapeutic modalities like small and large molecules, iPSC therapies pose significant unique challenges with respect to safety, potency, genetic stability, immunogenicity, tumorgenicity, cell reproducibility, scalability and engraftment. The aim of this review article is to highlight the unique technical challenges that need to be addressed before iPSC technology can be fully realized as a cell replacement therapy. Additionally, this manuscript offers some potential solutions and identifies areas of focus that should be considered in order for the iPSC field to achieve its promise. The scope of this article covers the following areas: (1) the impact of different iPSC reprogramming methods on immunogenicity and tumorigenicity; (2) the effect of genetic instability on cell reproducibility and differentiation; (3) the role of growth factors and post-translational modification on differentiation and cell scalability; (4) the potential use of gene editing in improving iPSC differentiation; (5) the advantages and disadvantages between autologous and allogeneic cell therapy; (6) the regulatory considerations in developing a viable and reproducible cell product; and (7) the impact of local tissue inflammation on cell engraftment and cell viability.
Masami Okamoto
Advanced Polymeric Nanostructured Materials Engineering, Graduate School of Engineering, Toyota Technological Institute, 2-12-1 Hisakata, Tempaku, Nagoya 468 8511, Japan
Abstract
Metastasis is one of the greatest challenges in cancer treatment today. Normal mammary epithelial cells are optimally supported by interaction with a soft matrix (microenvironment) with elastic modulus of about 800 Pa. However, after transformation, breast tissue becomes progressively stiffer and tumour cells become significantly more contractile and hyper-responsive to matrix elasticity. In addition, importantly, the cancer cells penetrate into blood vessel and enter the circulation during metastasis. The modulus of fluid such as blood or mucus has very low stiffness of around 50 Pa. For this reason, the critical association between cancer cell phenotype and the change of matrix rigidity with an order of magnitude smaller should be emphasizing. This review highlights the current understanding of epithelial-mesenchymal transition and cancer stem cells in metastasis, and identified importance for investigation on artificial extracellular matrix with different viscoelastic properties, which is required to mimics in vivo microenvironment. The substrate damping coefficient (tand) as potential physical parameter emerged the important linkage to cellular motility, cancer stemness, and epithelial-mesenchymal transition induction. Although further investigation is required to clarify the efficacy of environmental stimuli (tand) for tumors exhibiting stem cell-like properties, this review indicates that the cancer cells incubated on softer substrate might lead to express cancer stem cell biomarkers exhibiting high expression.
Aaron L. Sarver
Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455, USA; Institute for Health Informatics, University of Minnesota, Minneapolis, Minnesota 55455, USA; Animal Cancer Care and Research Program, University of Minnesota, St. Paul, Minnesota 55108, USA
Kelly M. Makielski
Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455, USA; Animal Cancer Care and Research Program, University of Minnesota, St. Paul, Minnesota 55108, USA; Department of Veterinary Clinical Sciences, University of Minnesota College of Veterinary Medicine, St. Paul, Minnesota 55108, USA
Jaime F. Modiano
Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455, USA; Animal Cancer Care and Research Program, University of Minnesota, St. Paul, Minnesota 55108, USA; Department of Veterinary Clinical Sciences, University of Minnesota College of Veterinary Medicine, St. Paul, Minnesota 55108, USA; Center for Immunology, University of Minnesota, Minneapolis, Minnesota 55455, USA; Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota 55455, USA; Center for Engineering and Medicine, University of Minnesota, Minneapolis, Minnesota 55455, USA; Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA
Abstract
This review will describe more than two decades of comparative research on primary bone cancer (osteosarcoma). Osteosarcoma is a chaotic disease present in a complex and variable microenvironment composed of many different cell types which interact with each other and lead to high transcriptional heterogeneity. Despite this heterogeneity, common transcriptional patterns can be observed in the bulk transcriptomes of these tumors; additionally, these patterns are associated with outcome, indicating their importance to the molecular biology of the disease. Work from our group and others has led to our current understanding of osteosarcoma as a disease where multiple pathological processes appear to converge into a limited array of tissue organizations with distinct biology. Recurrent as well as distinct events can lead to these states of tissue organization, explaining the heterogeneity of osteosarcoma that is observed among and within species. Yet, despite their chaotic genomes, osteosarcomas seem to be (relatively) genetically stable, with persistent maintenance of essentially the same chromothriptic karyotype throughout the developmental lifetime of the tumor. Importantly, the transcriptional variance between tumors can highlight the underlying biology of the malignant cells themselves, as well as the composition of the osteosarcoma microenvironment and the host response, both of which are prognostically significant for this disease. Initial single cell RNA-seq reports provide further evidence of the importance of the osteosarcoma microenvironment for tumor characterization. Our data suggest that improving patient outcomes in immunologically barren or “cold” osteosarcomas, necessitates generating immune permissive or “warmer” microenvironments within the tumor. Furthermore, the aging bone microenvironment may create specific niches that predispose to cancer, and identification of the drivers that lead to these variable transcriptional patterns will be essential to identify personalized, effective genomic therapy for osteosarcoma.
Farhad Nejat
Vision health research center, Tehran, Iran
Shima Eghtedari
Biomedical Engineering Department, Amirkabir University of Technology, Tehran, Iran
Abstract
Purpose: To introduce a novel technique using PANIS method (Plasma assisted noninvasive surgery) in pterygium removal surgery with conjunctival rotation flap.
Methods: Two patients (1 male, 1 Female) with grade 3 and 4 of pterygium, based on SLIT2 grading system, underwent surgery with local anesthesia, removing pterygium tissue from cornea and also all related subtenon. Then surgeon used residual conjunctiva to cover bare sclera using rotational method. Fusion of the border of conjunctival flaps from nasal to superior occurred by plasma spots. UCVA, BCVA, OSDI and patient satisfaction were evaluated before, 1 month, 6 months and 1 year after the surgery, as well as 1 day and 1 week post-operation examinations included slit-lamp and external check-up that has been done by the surgeon.
Results: Patients with primary pterygium, underwent surgery with PANIS method. Last follow-up session in 1 year, showed significant improvement in astigmatism and 1 line in BCVA. There were no complications and side effects during and after the surgery and also other reported complication included diplopia in lateral gaze, granuloma and scarring was not observed and no recurrence have been seen in the follow-up period and patients were satisfied. Additionally 1 day and 1 week slit-lamp examination showed plasma spots caused good attachment of conjunctival tissue with ocular surface.
Conclusion: After treating 2 patients with PANIS method, it seems that plasma spots can be a good substitute for suture or glue in pterygium surgery with conjunctival rotational flap, because it’s easy and fast way for fusing the conjunctival flap without any complication or side effect.
James E. Galen
Center for Vaccine Development and Global Health, Baltimore MD 21201; University of Maryland School of Medicine, Baltimore MD 21201
Thanh Pham
Center for Vaccine Development and Global Health, Baltimore MD 21201; University of Maryland School of Medicine, Baltimore MD 21201
Abstract
Cancer remains a serious challenge to public health, with breast cancer, lung, and colorectal cancers predominating in both incidence and deaths worldwide. Significant advances have been made in the therapeutic treatment and resolution of non-solid tumors using immune checkpoint inhibitors (ICI) and chimeric antigen receptor (CAR) T cell therapy to break immune tolerance and initiate tumor clearance. However, these innovative strategies have enjoyed only limited success with solid tumors, especially in late-stage cancers in which tumor size is large. An immunosuppressive tumor microenvironment (TME) that surrounds and protects solid tumors significantly confounds the ability of the host immune system to target and eliminate tumor tissue. Novel technologies using nanomedicines have begun to yield promising results by penetrating into the microenvironment to stimulate innate immunity and induce trafficking of activated antigen presenting cells to regional lymph nodes, ultimately leading to tumor-specific adaptive immune responses. One type of nanomedicine that is generating increasing enthusiasm in the field of immunotherapy are bacterial outer membrane vesicles (OMVs) that can be genetically engineered to surface-express tumor-associated antigens; the resulting recombinant OMVs (rOMVs) can then be purified as immunotherapeutic vaccines. Recent data from experimental animal models have demonstrated remarkable efficacy in tumor challenge models. Such promising experiments suggest the possibility of translating these novel strategies into success with solid tumors in clinical trials. In this review, we will summarize current research using purified rOMVs as immunotherapeutic vaccines and further discuss potential obstacles that still need to be adequately addressed to ensure success in human trials.
