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Ex vivo Culture of Fetal Mouse Gastric Epithelial Progenitors

Featured protocol,  Authors: Valeria Fernandez Vallone
Valeria Fernandez ValloneAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3957
Morgane Leprovots
Morgane LeprovotsAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3956
Gilbert Vassart
Gilbert VassartAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3958
 and Marie-Isabelle Garcia
Marie-Isabelle GarciaAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
For correspondence: mgarcia@ulb.ac.be
Bio-protocol author page: a3959
date: 1/5/2017, 91 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2089.

Brief version appeared in Development, May 2016
Isolation and tridimensional culture of murine fetal progenitors from the digestive tract represents a new approach to study the nature and the biological characteristics of these epithelial cells that are present before the onset of the cytodifferentiation process during development. In 2013, Mustata et al. described the isolation of intestinal fetal progenitors growing as spheroids in the ex vivo culture system initially implemented by Sato et al. (2009) to grow adult intestinal stem cells. Noteworthy, fetal-derived spheroids have high self-renewal capacity making easy their indefinite maintenance in culture. Here, we report an adapted protocol for isolation and ex vivo culture and maintenance of fetal epithelial progenitors from distal pre-glandular stomach growing as gastric spheroids (Fernandez Vallone et al., 2016).

Ex vivo Culture of Adult Mouse Antral Glands

Featured protocol,  Authors: Valeria Fernandez Vallone
Valeria Fernandez ValloneAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3957
Morgane Leprovots
Morgane LeprovotsAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3956
Gilbert Vassart
Gilbert VassartAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3958
 and Marie-Isabelle Garcia
Marie-Isabelle GarciaAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
For correspondence: mgarcia@ulb.ac.be
Bio-protocol author page: a3959
date: 1/5/2017, 81 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2088.

Brief version appeared in Development, May 2016
The tri-dimensional culture, initially described by Sato et al. (2009) in order to isolate and characterize epithelial stem cells of the adult small intestine, has been subsequently adapted to many different organs. One of the first examples was the isolation and culture of antral stem cells by Barker et al. (2010), who efficiently generated organoids that recapitulate the mature pyloric epithelium in vitro. This ex vivo approach is suitable and promising to study gastric function in homeostasis as well as in disease. We have adapted Barker’s protocol to compare homeostatic and regenerating tissues and here, we meticulously describe, step by step, the isolation and culture of antral glands as well as the isolation of single cells from antral glands that might be useful for culture after cell sorting as an example (Fernandez Vallone et al., 2016).

Gene Expression Analysis of Sorted Cells by RNA-seq in Drosophila Intestine

Featured protocol,  Authors: Jun Chen
Jun ChenAffiliation: National Institute of Biological Sciences, Beijing, China
Bio-protocol author page: a3929
Jia Li
Jia LiAffiliation: National Institute of Biological Sciences, Beijing, China
Bio-protocol author page: a16
Huaiwei Huang
Huaiwei HuangAffiliation: National Institute of Biological Sciences, Beijing, China
Bio-protocol author page: a3931
 and Rongwen Xi
Rongwen XiAffiliation: National Institute of Biological Sciences, Beijing, China
For correspondence: xirongwen@nibs.ac.cn
Bio-protocol author page: a3932
date: 12/20/2016, 185 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2079.

Brief version appeared in eLife, Jun 2016
RNA sequencing (RNA-seq) has become a popular method for profiling gene expression. Among many applications, one common purpose is to identify differentially expressed genes and pathways in different biological or pathological conditions. This protocol provides detailed procedure for RNA-seq analysis of ~250,000 sorted Drosophila intestinal cells (Chen et al., 2016), in which RNA amplification is not required.

Quantitative 3D Time Lapse Imaging of Muscle Progenitors in Skeletal Muscle of Live Mice

Featured protocol,  Authors: Micah T. Webster
Micah T. WebsterAffiliation: Department of Embryology, Carnegie Institution for Science, Baltimore, USA
Bio-protocol author page: a3892
Tyler Harvey
Tyler HarveyAffiliation 1: Department of Embryology, Carnegie Institution for Science, Baltimore, USA
Affiliation 2: Department of Biology, Johns Hopkins University, Baltimore, USA
Bio-protocol author page: a3893
 and Chen-Ming Fan
Chen-Ming FanAffiliation: Department of Embryology, Carnegie Institution for Science, Baltimore, USA
For correspondence: fan@ciwemb.edu
Bio-protocol author page: a3894
date: 12/20/2016, 133 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2066.

Brief version appeared in Cell Stem Cell, Feb 2016
For non-optically clear mammalian tissues, it is now possible to use multi-photon microscopy to penetrate deep into the tissue and obtain detailed single cell images in a live animal, i.e., intravital imaging. This technique is in principle applicable to any fluorescently marked cell, and we have employed it to observe stem cells during the regenerative process. Stem cell-mediated skeletal muscle regeneration in the mouse model has been classically studied at specific time points by sacrificing the animal and harvesting the muscle tissue for downstream analyses. A method for direct visualization of muscle stem cells to gain real-time information over a long period in a live mammal has been lacking. Here we describe a step-by-step protocol adapted from Webster et al. (2016) to quantitatively measure the behaviors of fluorescently labeled (GFP, EYFP) muscle stem and progenitor cells during homeostasis as well as following muscle injury.

Ex vivo Culture of Fetal Mouse Gastric Epithelial Progenitors

Authors: Valeria Fernandez Vallone
Valeria Fernandez ValloneAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3957
Morgane Leprovots
Morgane LeprovotsAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3956
Gilbert Vassart
Gilbert VassartAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3958
 and Marie-Isabelle Garcia
Marie-Isabelle GarciaAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
For correspondence: mgarcia@ulb.ac.be
Bio-protocol author page: a3959
date: 1/5/2017, 91 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2089.

[Abstract] Isolation and tridimensional culture of murine fetal progenitors from the digestive tract represents a new approach to study the nature and the biological characteristics of these epithelial cells that are present before the onset of the cytodifferentiation process during development. In 2013, Mustata et al. described the isolation of intestinal fetal ...

Ex vivo Culture of Adult Mouse Antral Glands

Authors: Valeria Fernandez Vallone
Valeria Fernandez ValloneAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3957
Morgane Leprovots
Morgane LeprovotsAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3956
Gilbert Vassart
Gilbert VassartAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
Bio-protocol author page: a3958
 and Marie-Isabelle Garcia
Marie-Isabelle GarciaAffiliation: Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium
For correspondence: mgarcia@ulb.ac.be
Bio-protocol author page: a3959
date: 1/5/2017, 81 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2088.

[Abstract] The tri-dimensional culture, initially described by Sato et al. (2009) in order to isolate and characterize epithelial stem cells of the adult small intestine, has been subsequently adapted to many different organs. One of the first examples was the isolation and culture of antral stem cells by Barker et al. (2010), who efficiently generated organoids ...

Gene Expression Analysis of Sorted Cells by RNA-seq in Drosophila Intestine

Authors: Jun Chen
Jun ChenAffiliation: National Institute of Biological Sciences, Beijing, China
Bio-protocol author page: a3929
Jia Li
Jia LiAffiliation: National Institute of Biological Sciences, Beijing, China
Bio-protocol author page: a16
Huaiwei Huang
Huaiwei HuangAffiliation: National Institute of Biological Sciences, Beijing, China
Bio-protocol author page: a3931
 and Rongwen Xi
Rongwen XiAffiliation: National Institute of Biological Sciences, Beijing, China
For correspondence: xirongwen@nibs.ac.cn
Bio-protocol author page: a3932
date: 12/20/2016, 185 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2079.

[Abstract] RNA sequencing (RNA-seq) has become a popular method for profiling gene expression. Among many applications, one common purpose is to identify differentially expressed genes and pathways in different biological or pathological conditions. This protocol provides detailed procedure for RNA-seq analysis of ~250,000 sorted Drosophila intestinal cells (Chen ...

Quantitative 3D Time Lapse Imaging of Muscle Progenitors in Skeletal Muscle of Live Mice

Authors: Micah T. Webster
Micah T. WebsterAffiliation: Department of Embryology, Carnegie Institution for Science, Baltimore, USA
Bio-protocol author page: a3892
Tyler Harvey
Tyler HarveyAffiliation 1: Department of Embryology, Carnegie Institution for Science, Baltimore, USA
Affiliation 2: Department of Biology, Johns Hopkins University, Baltimore, USA
Bio-protocol author page: a3893
 and Chen-Ming Fan
Chen-Ming FanAffiliation: Department of Embryology, Carnegie Institution for Science, Baltimore, USA
For correspondence: fan@ciwemb.edu
Bio-protocol author page: a3894
date: 12/20/2016, 133 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2066.

[Abstract] For non-optically clear mammalian tissues, it is now possible to use multi-photon microscopy to penetrate deep into the tissue and obtain detailed single cell images in a live animal, i.e., intravital imaging. This technique is in principle applicable to any fluorescently marked cell, and we have employed it to observe stem cells during the regenerative ...

In vitro Chondrogenic Hypertrophy Induction of Mesenchymal Stem Cells

Authors: Sang Young Jeong
Sang Young JeongAffiliation: Biomedical Research Institute, R&D Center, MEDIPOST Co., Ltd., Gyeonggi-do, Republic of Korea
Bio-protocol author page: a3855
Miyoung Lee
Miyoung LeeAffiliation: Biomedical Research Institute, R&D Center, MEDIPOST Co., Ltd., Gyeonggi-do, Republic of Korea
Bio-protocol author page: a3856
Soo Jin Choi
Soo Jin ChoiAffiliation: Biomedical Research Institute, R&D Center, MEDIPOST Co., Ltd., Gyeonggi-do, Republic of Korea
Bio-protocol author page: a3857
Wonil Oh
Wonil OhAffiliation: Biomedical Research Institute, R&D Center, MEDIPOST Co., Ltd., Gyeonggi-do, Republic of Korea
Bio-protocol author page: a3858
 and Hong Bae Jeon
Hong Bae JeonAffiliation: Biomedical Research Institute, R&D Center, MEDIPOST Co., Ltd., Gyeonggi-do, Republic of Korea
For correspondence: jhb@medi-post.co.kr
Bio-protocol author page: a3859
date: 12/5/2016, 229 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2057.

[Abstract] To investigate underlying mechanism of chondrogenic hypertrophy, we need proper in vitro hypertrophic model of mesenchymal stem cells (MSCs). This protocol describes our defined method for induction of in vitro chondrogenic hypertrophy of human umbilical cord blood-derived MSCs (hUCB-MSCs). By adding thyroid hormone (T3; triiodothyronine) and minimum ...

Isolation and Culture of Human Adipose-derived Stem Cells from Subcutaneous and Visceral White Adipose Tissue Compartments

Authors: Xiaojia Ge
Xiaojia GeAffiliation: Cell & Molecular Biology Group, Singapore Institute for Clinical Sciences (SICS), Agency for Science Technology and Research (A*STAR), Singapore, Singapore
Bio-protocol author page: a3759
Shi Chi Leow
Shi Chi LeowAffiliation: Cell & Molecular Biology Group, Singapore Institute for Clinical Sciences (SICS), Agency for Science Technology and Research (A*STAR), Singapore, Singapore
Bio-protocol author page: a3760
Durgalakshmi Sathiakumar
Durgalakshmi SathiakumarAffiliation: Cell & Molecular Biology Group, Singapore Institute for Clinical Sciences (SICS), Agency for Science Technology and Research (A*STAR), Singapore, Singapore
Bio-protocol author page: a3761
Walter Stünkel
Walter StünkelAffiliation: Cell & Molecular Biology Group, Singapore Institute for Clinical Sciences (SICS), Agency for Science Technology and Research (A*STAR), Singapore, Singapore
Bio-protocol author page: a3762
Asim Shabbir
Asim ShabbirAffiliation: Department of Surgery, National University Hospital, National University of Singapore, Singapore, Singapore
Bio-protocol author page: a3763
Jimmy Bok Yan So
Jimmy Bok Yan SoAffiliation: Department of Surgery, National University Hospital, National University of Singapore, Singapore, Singapore
Bio-protocol author page: a3764
Davide Lomanto
Davide LomantoAffiliation: Department of Surgery, National University Hospital, National University of Singapore, Singapore, Singapore
Bio-protocol author page: a3765
 and Craig McFarlane
Craig McFarlaneAffiliation: Cell & Molecular Biology Group, Singapore Institute for Clinical Sciences (SICS), Agency for Science Technology and Research (A*STAR), Singapore, Singapore
For correspondence: craig_mcfarlane@sics.a-star.edu.sg
Bio-protocol author page: a3766
date: 11/20/2016, 281 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2027.

[Abstract] Human Adipose-derived Stem/Stromal Cells (ASCs) have been widely used in stem cell and obesity research, as well as clinical applications including cell-based therapies, tissue engineering and reconstruction. Compared with mesenchymal stem cells (MSCs) derived from other tissues such as umbilical cord and bone marrow, isolation of ASCs from human white ...

Isolation and Primary Culture of Various Cell Types from Whole Human Endometrial Biopsies

Authors: Flavio Santos Vasconcelos Barros
Flavio Santos Vasconcelos BarrosAffiliation: Department of Biomedical Sciences, University of Warwick, Coventry, UK
Bio-protocol author page: a3767
Jan Joris Brosens
Jan Joris BrosensAffiliation: Department of Biomedical Sciences, University of Warwick, Coventry, UK
For correspondence: J.J.Brosens@warwick.ac.uk
Bio-protocol author page: a3768
 and Paul John Brighton
Paul John BrightonAffiliation: Department of Biomedical Sciences, University of Warwick, Coventry, UK
Bio-protocol author page: a3769
date: 11/20/2016, 246 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2028.

[Abstract] The isolation and primary culture of cells from human endometrial biopsies provides valuable experimental material for reproductive and gynaecological research. Whole endometrial biopsies are collected from consenting women and digested with collagenase and DNase I to dissociate cells from the extracellular matrix. Cell populations are then isolated ...

Allogeneic Transplantation of Testicular Hyperplasia in rag1 Mutant Zebrafish

Authors: Toshihiro Kawasaki
Toshihiro Kawasaki Affiliation: Genetic Strains Research Center, National Institute of Genetics, and Department of Genetics, School of Life Science, the Graduate University for Advanced Studies (SOKENDAI), Mishima, Japan
Bio-protocol author page: a3674
 and Noriyoshi Sakai
Noriyoshi SakaiAffiliation: Genetic Strains Research Center, National Institute of Genetics, and Department of Genetics, School of Life Science, the Graduate University for Advanced Studies (SOKENDAI), Mishima, Japan
For correspondence: nosakai@nig.ac.jp
Bio-protocol author page: a3675
date: 11/5/2016, 223 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1992.

[Abstract] Allogeneic organ transplantation is a powerful tool for clinical and basic research studies. However, the graft is often rejected by the host organism. Here, we describe a protocol that uses immunodeficient rag1 mutant zebrafish. These zebrafish escaped rejection, which made it possible to successfully transplant fragments of an allogeneic testis and ...

Experimental Liver Fibrosis and Intrasplenic Transplantation of CD45+ Bone Marrow Cells

Authors: Prakash Baligar
Prakash BaligarAffiliation: Amity Institute of Molecular Medicine and Stem Cell Research (AIMMSCR), Amity University, Noida, UP, India
For correspondence: pbaligar@amity.edu
Bio-protocol author page: a3620
Sebanta Pokhrel
Sebanta PokhrelAffiliation: Stem Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India
Bio-protocol author page: a3621
 and Asok Mukhopadhyay
Asok MukhopadhyayAffiliation: Stem Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India
For correspondence: ashok@nii.ac.in
Bio-protocol author page: a3622
date: 10/20/2016, 282 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1972.

[Abstract] Liver fibrosis results from the excessive collagen deposition (collagen scar) by activated hepatic stellate cells (HpSCs), leading to the inhibition of normal liver regeneration and function. Fibrogenesis is a complex mechanism involving both the synthesis and degradation of matrix proteins by different cell types, mainly macrophages in the liver. ...

Isolation, Culture, and Staining of Single Myofibers

Authors: Yann Simon Gallot
Yann Simon GallotAffiliation: Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA
Bio-protocol author page: a3535
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
Aman K. Mann
Aman K. MannAffiliation: duPont Manual High Schoo, Louisville, USA
Bio-protocol author page: a3538
 and Ashok Kumar
Ashok KumarAffiliation: 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: 10/5/2016, 461 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1942.

[Abstract] Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ...
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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, 10544 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, 8791 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, 8591 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, 7942 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, 7768 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 ...

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, 7167 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, 6704 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 ...

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, 6330 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 ...

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, 6277 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 ...

Isolation of Human Blood Progenitor and Stem Cells from Peripheral Blood by Magnetic Bead

Authors: Salma Hasan
Salma HasanAffiliation: INSERM U1009, Gustave Roussy, Villejuif, France
Bio-protocol author page: a140
 and Isabelle Plo
Isabelle PloAffiliation: INSERM U1009, Gustave Roussy, Villejuif, France
For correspondence: isabelle.plo@gustaveroussy.fr
Bio-protocol author page: a141
date: 11/5/2012, 5979 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.281.

[Abstract] The antigen CD34 is a well-known marker present on human progenitor and stem cells. This protocol explains the isolation of CD34+ cells from peripheral blood using magnetic bead separation technique. The approximate abundance of CD34+ cells in blood is 0.1% of mononuclear cells....
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