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Implantation of Human Peripheral Corneal Spheres into Cadaveric Human Corneal Tissue

Featured protocol,  Authors: Jeremy John Mathan
Jeremy John MathanAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
Bio-protocol author page: a4911
Salim Ismail
Salim IsmailAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
Bio-protocol author page: a4912
Jennifer Jane McGhee
Jennifer Jane McGheeAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
Bio-protocol author page: a4913
 and Trevor Sherwin
Trevor SherwinAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
For correspondence: t.sherwin@auckland.ac.nz
Bio-protocol author page: a4914
date: 7/20/2017, 14 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2412.

Brief version appeared in Stem Cell Res Ther, Jun 2016
Stem and progenitor cells isolated from human limbal tissue can be cultured in vitro as spheres. These spheres have potential for use as transplantable elements for the repopulation of corneal tissue (Mathan et al., 2016). Herein we describe the detailed protocol for the implantation of human corneal spheres into cadaveric human corneal tissue. This protocol describes the procedure for sphere formation and culture, preparation of tissue for sphere implantation, corneal limbus microsurgery and sphere implantation.

Microvesicle Isolation from Rat Brain Extract Treated Human Mesenchymal Stem Cells

Featured protocol,  Authors: Ji Yong Lee
Ji Yong LeeAffiliation: Institute for BioMedical Convergence, Catholic Kwandong University-International St. Mary’s Hospital, Incheon-si, Republic of Korea
Bio-protocol author page: a4768
Seong-Mi Choi
Seong-Mi ChoiAffiliation: Institute for BioMedical Convergence, Catholic Kwandong University-International St. Mary’s Hospital, Incheon-si, Republic of Korea
Bio-protocol author page: a4769
 and Han-Soo Kim
Han-Soo KimAffiliation: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea
For correspondence: hankim63@gmail.com
Bio-protocol author page: a4770
date: 7/5/2017, 212 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2375.

Brief version appeared in Sci Rep, Sep 2016
Microvesicle (MVs) are submicron-sized membranous vesicles that are either actively released from cells via secretory compartments or shed from cell surface membranes. MVs are generated by many cell types and serve as vehicles that transfer biological information (e.g., protein, mRNA, and miRNA) to distant cells, thereby affecting their gene expression, proliferation, differentiation, and function. Although their physiological functions are not clearly defined, recent studies have shown their therapeutic potential for tissue repair and regeneration. While MVs can be isolated readily from mesenchymal stem cells (MSCs) and other cell types from various sources, the yield of MVs under conventional culture condition in vitro is one of the limiting factors for both the in vivo functional study as well as in vitro molecular analysis. Here, we provide a protocol to increase the yield of microvesicles by preconditioning MSCs with rat brain extract.

Formaldehyde Fixation of Extracellular Matrix Protein Layers for Enhanced Primary Cell Growth

Featured protocol,  Authors: Natalia V. Andreeva
Natalia V. AndreevaAffiliation: Laboratory of Stem and Progenitor Cell Biology, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
Bio-protocol author page: a4766
 and Alexander V. Belyavsky
Alexander V. BelyavskyAffiliation: Laboratory of Stem and Progenitor Cell Biology, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
For correspondence: abelyavs@yahoo.com
Bio-protocol author page: a4767
date: 7/5/2017, 225 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2374.

Brief version appeared in Anal Biochem, Dec 2016
Coating tissue culture vessels with the components of the extracellular matrix such as fibronectin and collagens provides a more natural environment for primary cells in vitro and stimulates their proliferation. However, the effects of such protein layers are usually rather modest, which might be explained by the loss immobilized proteins due to their weak non-covalent association with the tissue culture plastic. Here we describe a simple protocol for a controlled fixation of fibronectin, vitronectin and collagen IV layers by formaldehyde, which substantially enhances the stimulation of primary cell proliferation by these extracellular proteins.

Implantation of Human Peripheral Corneal Spheres into Cadaveric Human Corneal Tissue

Authors: Jeremy John Mathan
Jeremy John MathanAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
Bio-protocol author page: a4911
Salim Ismail
Salim IsmailAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
Bio-protocol author page: a4912
Jennifer Jane McGhee
Jennifer Jane McGheeAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
Bio-protocol author page: a4913
 and Trevor Sherwin
Trevor SherwinAffiliation: Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, the University of Auckland, Private Bag 92019, Auckland, New Zealand
For correspondence: t.sherwin@auckland.ac.nz
Bio-protocol author page: a4914
date: 7/20/2017, 14 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2412.

[Abstract] Stem and progenitor cells isolated from human limbal tissue can be cultured in vitro as spheres. These spheres have potential for use as transplantable elements for the repopulation of corneal tissue (Mathan et al., 2016). Herein we describe the detailed protocol for the implantation of human corneal spheres into cadaveric human corneal tissue. This ...

Microvesicle Isolation from Rat Brain Extract Treated Human Mesenchymal Stem Cells

Authors: Ji Yong Lee
Ji Yong LeeAffiliation: Institute for BioMedical Convergence, Catholic Kwandong University-International St. Mary’s Hospital, Incheon-si, Republic of Korea
Bio-protocol author page: a4768
Seong-Mi Choi
Seong-Mi ChoiAffiliation: Institute for BioMedical Convergence, Catholic Kwandong University-International St. Mary’s Hospital, Incheon-si, Republic of Korea
Bio-protocol author page: a4769
 and Han-Soo Kim
Han-Soo KimAffiliation: Department of Biomedical Sciences, College of Medical Convergence, Catholic Kwandong University, Gangneung-si, Gangwon-do, Republic of Korea
For correspondence: hankim63@gmail.com
Bio-protocol author page: a4770
date: 7/5/2017, 212 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2375.

[Abstract] Microvesicle (MVs) are submicron-sized membranous vesicles that are either actively released from cells via secretory compartments or shed from cell surface membranes. MVs are generated by many cell types and serve as vehicles that transfer biological information (e.g., protein, mRNA, and miRNA) to distant cells, thereby affecting their gene expression, ...

Formaldehyde Fixation of Extracellular Matrix Protein Layers for Enhanced Primary Cell Growth

Authors: Natalia V. Andreeva
Natalia V. AndreevaAffiliation: Laboratory of Stem and Progenitor Cell Biology, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
Bio-protocol author page: a4766
 and Alexander V. Belyavsky
Alexander V. BelyavskyAffiliation: Laboratory of Stem and Progenitor Cell Biology, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
For correspondence: abelyavs@yahoo.com
Bio-protocol author page: a4767
date: 7/5/2017, 225 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2374.

[Abstract] Coating tissue culture vessels with the components of the extracellular matrix such as fibronectin and collagens provides a more natural environment for primary cells in vitro and stimulates their proliferation. However, the effects of such protein layers are usually rather modest, which might be explained by the loss immobilized proteins due to their ...

Transplantation of Embryonic Cortical Tissue into Lesioned Adult Brain in Mice

Authors: Cong Wang
Cong WangAffiliation: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou, China
Bio-protocol author page: a4734
Hao Gao
Hao GaoAffiliation: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou, China
Bio-protocol author page: a4735
 and Shengxiang Zhang
Shengxiang ZhangAffiliation: Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou, China
For correspondence: sxzhang@lzu.edu.cn
Bio-protocol author page: a4736
date: 6/20/2017, 238 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2360.

[Abstract] Transplantation of embryonic cortical tissue for repairing the damaged brain has provided a potential therapy for brain injury and diseases. The grafted tissue can successfully survive and participate in reestablishing the functional neural circuit of the host brain. Transplantation surgery can be combined with fluorescently labeled transgenic mice ...

Functional Analysis of Connexin Channels in Cultured Cells by Neurobiotin Injection and Visualization

Authors: Philipp Wörsdörfer
Philipp WörsdörferAffiliation: Institute of Anatomy and Cell Biology, University of Würzburg, Koellikerstraße 6, 97070 Würzburg, Germany
For correspondence: philipp.woersdoerfer@uni-wuerzburg.de
Bio-protocol author page: a4617
 and Klaus Willecke
Klaus WilleckeAffiliation: LIMES Institute, Molecular Genetics, University of Bonn, Carl-Troll-Straße 31, 53115 Bonn, Germany
Bio-protocol author page: a4618
date: 6/5/2017, 420 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2325.

[Abstract] Functional gap junction channels between neighboring cells can be assessed by microinjection of low molecular weight tracer substances into cultured cells. The extent of direct intercellular communication can be precisely quantified by this method. This protocol describes the iontophoretic injection and visualisation of Neurobiotin into cultured cells. ...

Efficient Production of Functional Human NKT Cells from Induced Pluripotent Stem Cells − Reprogramming of Human Vα24+iNKT Cells

Authors: Daisuke Yamada
Daisuke YamadaAffiliation: Laboratory for Developmental Genetics, Center for Integrative Medical Sciences (IMS), RIKEN, Yokohama, Japan
For correspondence: daisuke.yamada@riken.jp
Bio-protocol author page: a4503
Tomonori Iyoda
Tomonori IyodaAffiliation: Laboratory for Immunotherapy, Center for Integrative Medical Sciences (IMS), RIKEN, Yokohama, Japan
Bio-protocol author page: a4497
Kanako Shimizu
Kanako ShimizuAffiliation: Laboratory for Immunotherapy, Center for Integrative Medical Sciences (IMS), RIKEN, Yokohama, Japan
Bio-protocol author page: a4498
Yusuke Sato
Yusuke SatoAffiliation: Laboratory for Immunotherapy, Center for Integrative Medical Sciences (IMS), RIKEN, Yokohama, Japan
Bio-protocol author page: a4501
Haruhiko Koseki
Haruhiko KosekiAffiliation: Laboratory for Developmental Genetics, Center for Integrative Medical Sciences (IMS), RIKEN, Yokohama, Japan
Bio-protocol author page: a4502
 and Shin-ichiro Fujii
Shin-ichiro FujiiAffiliation: Laboratory for Immunotherapy, Center for Integrative Medical Sciences (IMS), RIKEN, Yokohama, Japan
For correspondence: shin-ichiro.fujii@riken.jp
Bio-protocol author page: a4504
date: 5/20/2017, 425 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2277.

[Abstract] Antigen-specific T cell-derived induced pluripotent stem cells (iPSCs) have been shown to re-differentiate into functional T cells and thus provide a potential source of T cells that could be useful for cancer immunotherapy. Human Vα24+ invariant natural killer T (Vα24+iNKT) cells are subset of T cells that are characterized by the expression of an ...

Isolation, Culturing, and Differentiation of Primary Myoblasts from Skeletal Muscle of Adult Mice

Authors: Lubna Hindi
Lubna HindiAffiliation: Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
Bio-protocol author page: a4417
Joseph D. McMillan
Joseph D. McMillanAffiliation: Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
Bio-protocol author page: a4418
Dil Afroze
Dil AfrozeAffiliation: Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
Bio-protocol author page: a4419
Sajedah M. Hindi
Sajedah M. HindiAffiliation: Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
Bio-protocol author page: a3536
 and Ashok Kumar
Ashok KumarAffiliation 1: Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
Affiliation 2: Professor and Distinguished University Scholar, Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
For correspondence: ashok.kumar@louisville.edu
Bio-protocol author page: a3537
date: 5/5/2017, 758 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2248.

[Abstract] Myogenesis is a multi-step process that leads to the formation of skeletal muscle during embryonic development and repair of injured myofibers. In this process, myoblasts are the main effector cell type which fuse with each other or to injured myofibers leading to the formation of new myofibers or regeneration of skeletal muscle in adults. Many steps ...

Analysis of Mitochondrial Transfer in Direct Co-cultures of Human Monocyte-derived Macrophages (MDM) and Mesenchymal Stem Cells (MSC)

Authors: Megan V. Jackson
Megan V. JacksonAffiliation: Centre for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, Belfast , Northern Ireland
Bio-protocol author page: a4447
 and Anna D. Krasnodembskaya
Anna D. KrasnodembskayaAffiliation: Centre for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, Belfast , Northern Ireland
For correspondence: a.krasnodembskaya@qub.ac.uk
Bio-protocol author page: a4449
date: 5/5/2017, 611 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2255.

[Abstract] Mesenchymal stem/stromal cells (MSC) are adult stem cells which have been shown to improve survival, enhance bacterial clearance and alleviate inflammation in pre-clinical models of acute respiratory distress syndrome (ARDS) and sepsis. These diseases are characterised by uncontrolled inflammation often underpinned by bacterial infection. The mechanisms ...

Explant Methodology for Analyzing Neuroblast Migration

Authors: Kirsty J. Dixon
Kirsty J. DixonAffiliation: Department of Transplant Surgery, Virginia Commonwealth University, Richmond, USA
Bio-protocol author page: a4427
Alisa Turbic
Alisa TurbicAffiliation: Department of Epidemiology and Preventive Medicine, Monsah University, Melbourne, Australia
Bio-protocol author page: a4428
Ann M. Turnley
Ann M. TurnleyAffiliation: Department of Anatomy and Neuroscience, The University of Melbourne, Parkville, Australia
Bio-protocol author page: a4429
 and Daniel J. Liebl
Daniel J. LieblAffiliation: The Miami Project to Cure Paralysis and Department of Neurological Surgery, University of Miami, Miami, USA
For correspondence: dliebl@miami.edu
Bio-protocol author page: a4430
date: 5/5/2017, 457 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2249.

[Abstract] The subventricular zone (SVZ) in the mammalian forebrain contains stem/progenitor cells that migrate through the rostral migratory stream (RMS) to the olfactory bulb throughout adulthood. SVZ-derived explant cultures provide a convenient method to assess factors regulating the intermediary stage of neural stem/progenitor cell migration. Here, we describe ...

Mimicking Angiogenesis in vitro: Three-dimensional Co-culture of Vascular Endothelial Cells and Perivascular Cells in Collagen Type I Gels

Authors: Markus Auler
Markus AulerAffiliation 1: Medical Faculty, Department of Pediatrics and Adolescent Medicine, Experimental Neonatology, Cologne, Germany
Affiliation 2: Medical Faculty, Center for Biochemistry, University of Cologne, Cologne, Germany
Bio-protocol author page: a4408
Lena Pitzler
Lena PitzlerAffiliation 1: Medical Faculty, Department of Pediatrics and Adolescent Medicine, Experimental Neonatology, Cologne, Germany
Affiliation 2: Medical Faculty, Center for Biochemistry, University of Cologne, Cologne, Germany
Bio-protocol author page: a4409
Ernst Pöschl
Ernst PöschlAffiliation: Biomedical Research Centre, School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich, UK
Bio-protocol author page: a4410
Zhigang Zhou
Zhigang ZhouAffiliation 1: Biomedical Research Centre, School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich, UK
Affiliation 2: Medical Faculty, University of East Anglia, Norwich Research Park, Norwich, UK
Bio-protocol author page: a4411
 and Bent Brachvogel
Bent BrachvogelAffiliation 1: Medical Faculty, Department of Pediatrics and Adolescent Medicine, Experimental Neonatology, Cologne, Germany
Affiliation 2: Medical Faculty, Center for Biochemistry, University of Cologne, Cologne, Germany
For correspondence: bent.brachvogel@uni-koeln.de
Bio-protocol author page: a4412
date: 4/20/2017, 621 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2247.

[Abstract] Angiogenesis defines the process of formation of new vascular structures form existing blood vessels, involved during development, repair processes like wound healing but also linked to pathological changes. During angiogenic processes, endothelial cells build a vascular network and recruit perivascular cells to form mature, stable vessels. Endothelial ...
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Synchronize Human Embryonic Stem Cells at Different Cell Cycle Stage

Author: Hui Zhu
Hui ZhuAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: huizhu@stanford.edu
Bio-protocol author page: a32
date: 6/5/2012, 11904 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.193.

[Abstract] Pluripotency and the capability for self-renewal are essential characteristics of human embryonic stem cells (hESCs), which hold great potential as a cellular source for tissue replacement. Short cell cycle (15-16 h) compared to somatic cells is another property of hESCs. Efficient synchronization of ...

Generation of Mouse Bone Marrow-Derived Dendritic Cells (BM-DCs)

Authors: Francesca Granucci
Francesca GranucciAffiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
For correspondence: francesca.granucci@unimib.it
Bio-protocol author page: a59
Renato Ostuni
Renato OstuniAffiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
Bio-protocol author page: a58
 and Ivan Zanoni
Ivan ZanoniAffiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
For correspondence: ivan.zanoni@unimib.it
Bio-protocol author page: a54
date: 6/20/2012, 10043 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.226.

[Abstract] Generating mouse dendritic cells from bone-marrow progenitor cells is a useful tool to study biological functions of mouse dendritic cells. Dendritic cells are one of the major populations of phagocytes able to activate both innate and adaptive immune cells. ...

Sphere Formation (Osteosphere/Sarcopshere) Assay

Authors: Upal Basu-Roy
Upal Basu-RoyAffiliation: Department of Microbiology, NYU School of Medicine, New York, USA
For correspondence: upal.basuroy@nyumc.org
Bio-protocol author page: a190
Claudio Basilico
Claudio BasilicoAffiliation: Department of Microbiology, NYU School of Medicine, New York, USA
Bio-protocol author page: a191
 and Alka Mansukhani
Alka MansukhaniAffiliation: Department of Microbiology, NYU School of Medicine, New York, USA
For correspondence: alka.mansukhani@med.nyu.edu
Bio-protocol author page: a192
date: 12/20/2012, 9888 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.307.

[Abstract] Self-renewing cells from adult tissue (such as bone) that represent a progenitor population can be grown in suspension cultures in the presence of defined serum-free medium. Progenitor cells can be identified by this property of anchorage-independent growth in suspension cultures. These spherical clusters ...

[Bio101] Mouse Embryonic Stem Cell Maintenance for Differentiation

Author: Hogune Im
Hogune ImAffiliation: Molecular and Cellular Pharmacology Program, Department of Pharmacology, University of Wisconsin Medical Schoo, Madison, USA
For correspondence: hoguneim@stanford.edu
Bio-protocol author page: a18
date: 10/20/2011, 9129 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.145.

[Abstract] Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a protocol to maintain mouse stem cells ...

[Bio101] Culturing of C57BL/6 Mouse Embryonic Stem (ES) Cell Line

Author: Yuqiong Pan date: 10/5/2011, 8806 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.142.

[Abstract] Using ATCC ES C57BL/6 as an example, it is shown here how to culture mouse embryonic stem (ES) cell line. The clonal embryonic stem cell line #693 ES C57BL/6 was derived from a strain C57BL/6J (B6) mouse blastocyst [PubMed: 11730008]. The ES cells were shown to populate the germ line of two host blastocyst ...

Wnt Reporter Activity Assay

Author: Chen Zhao
Chen ZhaoAffiliation: Department of Developmental Biology, Institute for Stem Cell and Regenerative Medicine, Stanford University, Stanford, USA
For correspondence: chenzhao@stanford.edu
Bio-protocol author page: a1513
date: 7/20/2014, 8568 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1183.

[Abstract] This protocol is for testing responses of a candidate cell line/cell lines to Wnt ligands or Wnt pathway agonists stimulation. This protocol can also be adapted to screen small molecule libraries or biologics that contain activities to either increase or decrease Wnt pathway responses. Canonical Wnt ...

Mouse Embryonic Stem Cell Differentiation to Hematopoietic Precursors

Author: Hogune Im
Hogune ImAffiliation: Department of Pharmacology, University of Wisconsin Medical School, Madison, USA
For correspondence: hoguneim@stanford.edu
Bio-protocol author page: a18
date: 4/5/2012, 8189 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.144.

[Abstract] Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a two step protocol to form embryoid ...

Isolation and 3-dimensional Culture of Primary Murine Intestinal Epithelial Cells

Authors: Agnieszka Pastuła
Agnieszka PastułaAffiliation: II. Medizinische Klinik, Klinikum rechts der Isar, Technische Universität München, Munich, Germany
For correspondence: agnieszka.pastula@tum.de
Bio-protocol author page: a1359
 and  Michael Quante
Michael QuanteAffiliation: II. Medizinische Klinik, Klinikum rechts der Isar, Technische Universität München, Munich, Germany
Bio-protocol author page: a1360
date: 5/20/2014, 8179 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1125.

[Abstract] The intestine, together with skin and blood, belongs to the organs with the highest cell turnover, which makes it a perfect model to study cellular processes, such as proliferation and differentiation. Epithelial cell turnover in intestine is possible due to the presence of intestinal stem cells, which ...

Alkaline Phosphatase Staining

Author: Pearl A. Campbell
Pearl A. CampbellAffiliation: Sprott Centre for Stem Cell Research, Ottawa Hospital Research Institute, Ottawa, Canada
For correspondence: pcampbell@ohri.ca
Bio-protocol author page: a1193
date: 3/5/2014, 7909 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1060.

[Abstract] Two main features characterize pluripotent cells; self-renewal (unlimited cell division) and the ability to give rise to all cells of the adult organism. Given the recent impact of induced pluripotent stem cells (iPSCs) and ongoing use of pluripotent embryonic stem cells ESCs (ESCs) in basic discovery, ...

Single-cell Gene Expression Profiling of Mouse Stem Cells With Fluidigm BiomarkTM Dynamic Array

Author: Ana Sevilla
Ana SevillaAffiliation: Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine, New York, USA
For correspondence: ana.sevilla7@gmail.com
Bio-protocol author page: a351
date: 5/5/2013, 7649 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.692.

[Abstract] This protocol describes how to use Fluidigm BiomarkTM 96.96 dynamic arrays for high-throughput expression profiling from single mouse stem cells, assaying up to 96 independent samples with up to 96 quantitative PCR (qPCR) probes (equivalent to 9,216 reactions) in a single experiment. This Dynamic Array ...
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