Core Concepts in Stem Cell Biology: 1 Syllabus and Learning Guide
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Corneal Cell Therapy: with Ipscs, It Is No More a Far-Sight Koushik Chakrabarty1* , Rohit Shetty2 and Arkasubhra Ghosh1
Chakrabarty et al. Stem Cell Research & Therapy (2018) 9:287 https://doi.org/10.1186/s13287-018-1036-5 REVIEW Open Access Corneal cell therapy: with iPSCs, it is no more a far-sight Koushik Chakrabarty1* , Rohit Shetty2 and Arkasubhra Ghosh1 Abstract Human-induced pluripotent stem cells (hiPSCs) provide a personalized approach to study conditions and diseases including those of the eye that lack appropriate animal models to facilitate the development of novel therapeutics. Corneal disease is one of the most common causes of blindness. Hence, significant efforts are made to develop novel therapeutic approaches including stem cell-derived strategies to replace the diseased or damaged corneal tissues, thus restoring the vision. The use of adult limbal stem cells in the management of corneal conditions has been clinically successful. However, its limited availability and phenotypic plasticity necessitate the need for alternative stem cell sources to manage corneal conditions. Mesenchymal and embryonic stem cell-based approaches are being explored; nevertheless, their limited differentiation potential and ethical concerns have posed a significant hurdle in its clinical use. hiPSCs have emerged to fill these technical and ethical gaps to render clinical utility. In this review, we discuss and summarize protocols that have been devised so far to direct differentiation of human pluripotent stem cells (hPSCs) to different corneal cell phenotypes. With the summarization, our review intends to facilitate an understanding which would allow developing efficient and robust protocols to obtain specific corneal cell phenotype from hPSCs for corneal disease modeling and for the clinics to treat corneal diseases and injury. Keywords: Cornea, Induced pluripotent stem cells, Differentiation, Disease modeling, Cell replacement therapy Background using viral vectors. -
Selective Expansion of Limbal Epithelial Stem Cells in Culture Using Hypoxia
1 Selective Expansion of Limbal Epithelial Stem Cells in Culture Using Hypoxia Chris Bath1,2, Sufang Yang2,3, Danson Muttuvelu1, Trine Fink2, Jeppe Emmersen2, Henrik Vorum1, Jesper Hjortdal4 and Vladimir Zachar2 1Department of Ophthalmology, Aalborg University Hospital, 9000 Denmark 2Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220 Aalborg, Denmark 3Animal Reproduction Institute, Guangxi University, China 4Department of Ophthalmology, Aarhus University Hospital, 8000 Aarhus, Denmark Abstract Limbal epithelial stem cells (LESCs) maintain the corneal epithelium through- out life and are crucial for both corneal integrity and vision. In this study, LESCs were expanded in either a culture system using 3T3 feeder cells in growth medium supplemented with serum, or in a culture system without feeder cells using commercially available serum-free medium (EpiLife). Cells were maintained at an ambient oxygen concentration of 20% or at various levels of hypoxia (15%, 10%, 5%, and 2%) throughout the period of expansion. The effect of ambient oxygen concentration on growth, cell cycle, colony forming efficiency (CFE), and expression of stem cell markers ABCG2 and p63α and differentiation marker CK3 were determined at different time points. Low oxygen levels were found to maintain a stem cell phenotype with low proliferative rate, high CFE, and high expression of ABCG2 and p63α as well as low expression of CK3. The relation between degree of differentiation and ambient oxygen concentration in the culture system seems to mirror the natural environment of the limbal niche. Hypoxic culture could therefore potentially improve stem cell grafts for cultured limbal epithelial transplantation (CLET). 1 2 Selective Expansion of Limbal Epithelial Stem Cells in Culture Using Hypoxia Keywords: Limbus cornea, Adult stem cells, Regenerative medicine, Cell hypoxia, Primary cell culture. -
Bone Marrow (Stem Cell) Transplant for Sickle Cell Disease Bone Marrow (Stem Cell) Transplant
Bone Marrow (Stem Cell) Transplant for Sickle Cell Disease Bone Marrow (Stem Cell) Transplant for Sickle Cell Disease 1 Produced by St. Jude Children’s Research Hospital Departments of Hematology, Patient Education, and Biomedical Communications. Funds were provided by St. Jude Children’s Research Hospital, ALSAC, and a grant from the Plough Foundation. This document is not intended to take the place of the care and attention of your personal physician. Our goal is to promote active participation in your care and treatment by providing information and education. Questions about individual health concerns or specifi c treatment options should be discussed with your physician. For more general information on sickle cell disease, please visit our Web site at www.stjude.org/sicklecell. Copyright © 2009 St. Jude Children’s Research Hospital How did bone marrow (stem cell) transplants begin for children with sickle cell disease? Bone marrow (stem cell) transplants have been used for the treatment and cure of a variety of cancers, immune system diseases, and blood diseases for many years. Doctors in the United States and other countries have developed studies to treat children who have severe sickle cell disease with bone marrow (stem cell) transplants. How does a bone marrow (stem cell) transplant work? 2 In a person with sickle cell disease, the bone marrow produces red blood cells that contain hemoglobin S. This leads to the complications of sickle cell disease. • To prepare for a bone marrow (stem cell) transplant, strong medicines, called chemotherapy, are used to weaken or destroy the patient’s own bone marrow, stem cells, and infection fi ghting system. -
Induced Pluripotent Stem Cells As a Potential Therapeutic Source for Corneal Epithelial Stem Cells
iPSCs for corneal diseases ·Review· Induced pluripotent stem cells as a potential therapeutic source for corneal epithelial stem cells Jie Zhu1, Mark Slevin2,3, Bao-Qiang Guo2,3, Shou-Rong Zhu4 1Queen Mary School, Medical College of Nanchang Citation: Zhu J, Slevin M, Guo BQ, Zhu SR. Induced pluripotent University, Nanchang 330006, Jiangxi Province, China stem cells as a potential therapeutic source for corneal epithelial stem 2School of Healthcare Science, Faculty of Science and cells. Int J Ophthalmol 2018;11(12):2004-2010 Engineering, Manchester Metropolitan University, Chester Street, Manchester M15GD, United Kingdom INTRODUCTION 3Research Institute of Brain Vascular Disease, Weifang Medical luripotent stem cells are primitive cells that are able University, Weifang 261000, Shandong Province, China P to be self-renewing, proliferating indefinitely in their 4Department of Ophthalmology, Affiliated Hospital of Weifang undifferentiated state, and differentiate into different cell [1-2] Medical University, Weifang 261000, Shandong Province, types . They can efficiently differentiate into specific cell China types under defined conditions. Therefore, stem cells have Correspondence to: Bao-Qiang Guo. Research Institute of been regarded as unlimited source of cell transplantation. Brain Vascular Disease, Weifang Medical University, Weifang Over the last decade, stem or progenitor cells transplantation 261000, Shandong Province, China. [email protected]; as a means of replacing tissue have evolved rapidly. There are Shou-Rong Zhu. Department of Ophthalmology, Affiliated distinct kinds of stem cells according to their differentiation Hospital of Weifang Medical University, Weifang 261000, potential. It has already been found that embryonic stem cells Shandong Province, China. [email protected]. (ESCs) and mesenchymal stem cells (MSCs) can directly Received: 2018-08-21 Accepted: 2018-10-12 differentiate into specialized cells, which attracted considerable interest. -
Genetic Manipulation of Stem Cells Eleni Papanikolaou1,2*, Kalliopi I
logy & Ob o st ec e tr n i y c s G Papanikolaou et al. Gynecol Obstetric 2011, S:6 Gynecology & Obstetrics DOI: 10.4172/2161-0932.S6-001 ISSN: 2161-0932 Review Article Open Access Genetic Manipulation of Stem Cells Eleni Papanikolaou1,2*, Kalliopi I. Pappa1,3 and Nicholas P. Anagnou1,2 1Laboratory of Cell and Gene Therapy, Centre for Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece 2Laboratory of Biology, University of Athens School of Medicine, Athens, Greece 3First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, Athens, Greece Abstract Stem cells have the remarkable potential for self-renewal and differentiation into many cell types in the body during early life and development. In addition, in many tissues they constitute a source of internal repair system, dividing essentially without limit to replenish damaged or dead cells. After division, each new cell has the potential either to retain the stem cell status or to differentiate to a more specialized cell type, such as a red blood cell, a brain cell or a heart cell. Until recently, three types of stem cells from animals and humans have been characterized, i.e. embryonic stem cells, fetal stem cells and somatic adult stem cells. However, in late 2007, researchers accomplished another breakthrough by identifying conditions that allow some specialized adult cells to be “reprogrammed” genetically to assume a stem cell-like state. These cells, called induced pluripotent stem cells (iPSCs), express genes and factors important for maintaining the unique properties and features of embryonic stem cells. -
Properties of Mouse Inner Cell Masses Isolated by Immunosurgery, Exposure to the Calcium Ionophore A23187 Or Low Osmolarity
Aust. J. BioI. Sci., 1980,33, 689-97 Properties of Mouse Inner Cell Masses Isolated by Immunosurgery, Exposure to the Calcium Ionophore A23187 or Low Osmolarity G. M. Harlow and P. Quinn Department of Biological Sciences, University of Newcastle, Newcastle, N.S.W. 2308. Abstract Mouse inner cell masses remained intact when exposed to extreme bsmotic stress (distilled water) for short periods, but the trophectoderm was lysed in one-third of blastocysts. However, these inner cell masses were not viable as they could not fluoresce after incubation in fluorescein diacetate nor continue development in vitro. It was concluded that cells of the inner cell mass are not more tolerant of osmotic stresses than those of the trophectoderm. Inner cell masses were isolated from only 50 % of blastocysts when exposed to the calcium ionophore A23187. Some lysing trophectoderm cells are probably able to restore their normal calcium levels after transfer to fresh culture medium allowing normal blastocyst-like development in vitro. Further evidence which suggests that not all trophectoderm cells are removed after incu bation in ionophore is that these inner cell masses produced more trophoblast-like outgrowths in vitro and were able to induce the decidual cell reaction in vivo more often than their immuno surgically isolated counterparts. Irnmunosurgically isolated inner cell masses were viable as they fluoresced brightly after incu bation in fluorescein diacetate and developed normally in vitro. There was little or no contamination of these inner cell masses with trophectoderm cells as they formed considerably smaller outgrowths than control blastocysts and were rarely able to induce the decidual cell reaction in vivo. -
Genetics and Stem Cell Research A.Genetics
7: Genetics and Stem Cell Research A.Genetics 1. Introduction The principal special feature of genetics research is that the result of the study applies not only to the proband but also influences her lineage both in the past and in the future. For example genetic studies demonstrated Thomas Jefferson’s sexual relationship with his slave Sally Hemings and defined their descendants to this day. As we all know from television, genetic studies can be done from any tissue fragment that contains DNA so that studies of surgical specimens, biopsy materials, hair, epithelium and blood samples can all be utilized for extensive genetic studies. 2. Sampling Some DNA is more medically valuable than other. Samples from isolated populations in which a particular disorder is prevalent have a much greater probability of yielding the causal gene(s) because they have fewer genome variations than in the general population. Once isolated, the genetic material associated with the disorder has a good chance of yielding novel diagnostic and/or therapeutic approaches for the disorder. 3. Property rights A persistent question is whether the providers of the genetic material have any rights to the products created from their genetic material. These days, most consent forms are written explicitly to exclude intellectual property rights from the subjects. As might be imagined, this smacks of exploitation in the developing world. Negotiation of a monetary return to the community has sometimes been concluded. Important and lucrative products have been derived from individuals’ genomes without their receiving royalties or other compensation. However, the knowledge, technical expertise, and capital needed to make a useful product from a blood or tissue sample come from the company not the donor. -
The Amazing Stem Cell What Are They? Where Do They Come From? How Are They Changing Medicine? Stem Cells Are “Master Cells”
The Amazing Stem Cell What are they? Where do they come from? How are they changing medicine? Stem cells are “master cells” Stem cells can be “guided” to become many other cell types. Stem Cell Bone cell Self-renewed stem cell Brain cell Heart muscle Blood cell cell There are several types of stem cells, each from a unique source Embryonic stem cells* • Removed from embryos created for in vitro fertilization after donation consent is given. (Not sourced from aborted fetuses.) • Embryos are 3-5 days old (blastocyst) and have about 150 cells. • Can become any type of cell in the body, also called pluripotent cells. • Can regenerate or repair diseased tissue and organs. • Current use limited to eye-related disorders. * Not used by Mayo Clinic. Adult stem cells • Found in most adult organs and tissues, including bone marrow. • Often taken from bone marrow in the hip. • Blood stem cells can be collected through apheresis (separated from blood). • Can regenerate and repair diseased or damaged tissues (regenerative medicine). • Can be used as specialized “drugs” to potentially treat degenerative conditions. • Currently tested in people with neurological and heart disease. Umbilical cord blood stem cells • Found in blood in placenta and umbilical cord after childbirth. • Have the ability to change into specialized cells (like blood cells), also called progenitor cells. • Parents choose to donate umbilical cord blood for use in research, or have it stored for private or public banks. • Can be used in place of bone marrow stem cell transplants in some clinical applications. Bioengineered stem cells • Regular adult cells (e.g., blood, skin) reprogrammed to act like embryonic stem cells (induced pluripotent stem cells). -
The Longest Telomeres: a General Signature of Adult Stem Cell Compartments
Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press The longest telomeres: a general signature of adult stem cell compartments Ignacio Flores,1 Andres Canela,1 Elsa Vera,1 Agueda Tejera,1 George Cotsarelis,2 and María A. Blasco1,3 1Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre (CNIO), Madrid E-28029, Spain; 2University of Pennsylvania School of Medicine, M8 Stellar-Chance Laboratories, Philadelphia, Pennsylvania 19104, USA Identification of adult stem cells and their location (niches) is of great relevance for regenerative medicine. However, stem cell niches are still poorly defined in most adult tissues. Here, we show that the longest telomeres are a general feature of adult stem cell compartments. Using confocal telomere quantitative fluorescence in situ hybridization (telomapping), we find gradients of telomere length within tissues, with the longest telomeres mapping to the known stem cell compartments. In mouse hair follicles, we show that cells with the longest telomeres map to the known stem cell compartments, colocalize with stem cell markers, and behave as stem cells upon treatment with mitogenic stimuli. Using K15-EGFP reporter mice, which mark hair follicle stem cells, we show that GFP-positive cells have the longest telomeres. The stem cell compartments in small intestine, testis, cornea, and brain of the mouse are also enriched in cells with the longest telomeres. This constitutes the description of a novel general property of adult stem cell compartments. Finally, we make the novel finding that telomeres shorten with age in different mouse stem cell compartments, which parallels a decline in stem cell functionality, suggesting that telomere loss may contribute to stem cell dysfunction with age. -
Nano-Biosensor for Monitoring the Neural Differentiation of Stem Cells
nanomaterials Review Nano-Biosensor for Monitoring the Neural Differentiation of Stem Cells Jin-Ho Lee 1,2,†, Taek Lee 1,2,† and Jeong-Woo Choi 1,2,* 1 Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742, Korea; [email protected] (J.-H.L.); [email protected] (T.L.) 2 Institute of Integrated Biotechnology, Sogang University, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742, Korea * Correspondence: [email protected]; Tel.: +82-2-718-1976; Fax: +82-2-3273-0331 † These authors contributed equally to this work. Academic Editors: Chen-Zhong Li and Ling-Jie Meng Received: 6 July 2016; Accepted: 17 November 2016; Published: 28 November 2016 Abstract: In tissue engineering and regenerative medicine, monitoring the status of stem cell differentiation is crucial to verify therapeutic efficacy and optimize treatment procedures. However, traditional methods, such as cell staining and sorting, are labor-intensive and may damage the cells. Therefore, the development of noninvasive methods to monitor the differentiation status in situ is highly desirable and can be of great benefit to stem cell-based therapies. Toward this end, nanotechnology has been applied to develop highly-sensitive biosensors to noninvasively monitor the neural differentiation of stem cells. Herein, this article reviews the development of noninvasive nano-biosensor systems to monitor the neural differentiation of stem cells, mainly focusing on optical (plasmonic) and eletrochemical methods. The findings in this review suggest that novel nano-biosensors capable of monitoring stem cell differentiation are a promising type of technology that can accelerate the development of stem cell therapies, including regenerative medicine. -
Isolation of a Primate Embryonic Stem Cell Line JAMES A
Proc. Natl. Acad. Sci. USA Vol. 92, pp. 7844-7848, August 1995 Developmental Biology Isolation of a primate embryonic stem cell line JAMES A. THOMSON*t, JENNIFER KALISHMAN*, THADDEUS G. GOLOS*, MAUREEN DURNING*, CHARLEs P. HARRIS*, ROBERT A. BECKER*, AND JOHN P. HEARN*§ *The Wisconsin Regional Primate Research Center, §Department of Physiology, School of Medicine, and tCytogenetics Laboratory, State Hygiene Laboratory, University of Wisconsin, 1223 Capitol Court, Madison, WI 53715-1299 Communicated by Neal L. First, University of Wsconsin, Madison, W1, May 4, 1995 (received for review January 23, 1995) ABSTRACT Embryonic stem cells have the ability to fetal membranes and placenta (15), and in formation of an remain undifferentiated and proliferate indefinitely in vitro embryonic disc instead of an egg cylinder. Human embryonal while maintaining the potential to differentiate into deriva- carcinoma (EC) cells, which are pluripotent, immortal stem tives of all three embryonic germ layers. Here we report the cells from teratocarcinomas, provide an important in vitro derivation ofa cloned cell line (R278.5) from a rhesus monkey model for understanding human differentiation (16). Some EC blastocyst that remains undifferentiated in continuous pas- cell lines can be induced to differentiate in culture (17), which sage for >1 year, maintains a normal XY karyotype, and results in the loss ofspecific cell surface markers [stage-specific expresses the cell surface markers (alkaline phosphatase, embryonic antigen 3 (SSEA-3), SSEA-4, TRA-1-60, and TRA- stage-specific embryonic antigen 3, stage-specific embryonic 1-81] and the appearance of new markers (16). When pluri- antigen 4, TRA-1-60, and TRA-1-81) that are characteristic of potent human EC cells are injected into immunocompromised human embryonal carcinoma cells. -
Simple Limbal Epithelial Transplantation Method in the Treatment of Unilateral Limbal Stem Cell Deficiency Due to Chemical Burn
DOI: 10.14744/eer.2021.32032 Eur Eye Res 2021;1(1):31–36 ORIGINAL ARTICLE Simple limbal epithelial transplantation method in the treatment of unilateral limbal stem cell deficiency due to chemical burn Dilay Ozek, Emine Esra Karaca, Ozlem Evren Kemer Department of Opthalmology, Ankara City Hospital, Ankara, Turkey Abstract Purpose: The objectives of the study were to evaluate the success of the simple limbal epithelial transplantation (SLET) method in the treatment of unilateral limbal stem cell deficiency (LSCD) due to chemical burn. Methods: Seventeen patients with unilateral LSCD due to chemical burn were included in this retrospective study. Mean age of patients was 50.3±20.8 (28–75) years. Mean duration of follow was 18.9±6.9 (12–24) months. In the recipient eye follow- ing peritomy, pannus tissue was cleared and covered with amniotic membrane with fibrin glue. Limbal stem cell received from the fellow eye was implanted cornea surface 2–3 mm inside limbus with fibrin glue on the amniotic membrane and placed contact lens. In control examination of all patients who completed minimum 12 months postoperatively, regression in corneal vascularization, duration of epithelial healing, visual acuity, need for keratoplasty, and complications (dropping of contact lenses, separation of amniotic membrane, and graft failure) were evaluated. Results: Corneal epithelization was completed between 4 and 6 weeks in all patients. Total and partial separations in the amniotic membrane occurred in two patients. Marked regression in corneal vascularization and increase in visual acuity was observed in all patients. Five patients (29.4%) underwent keratoplasty in the follow-up period.