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In vitro Chondrogenic Hypertrophy Induction of Mesenchymal Stem Cells

Featured protocol,  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, 42 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2057.

Brief version appeared in Stem Cells, Nov 2015
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 osteogenic-inducing factors to culture medium, we could induce hypertrophy of hUCB-MSCs in vitro. Hypertrophic induction was validated using immunohistochemical analysis, Western blotting and reverse transcriptase polymerase chain reaction.

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

Featured protocol,  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, 116 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2027.

Brief version appeared in Eukaryot Cell, Oct 2007
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 adipose tissue (WAT) has great advantages due to its rich tissue source and simple surgical procedure. In this detailed protocol we describe a protocol to isolate and characterize ASCs from human WAT. Molecular characterization of isolated ASCs was performed through surface marker expression profiling using flow cytometry. Adipogenic capacity of the isolated ASCs was confirmed through inducing adipogenic differentiation and Oil Red O staining of lipid. This protocol provides researchers with the tools to culture and assess purity and adipogenic differentiation capacity of human ASCs, which can then be utilized for required downstream in vitro applications.

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

Featured protocol,  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, 102 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2028.

Brief version appeared in Stem Cells, Jul 2015
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 through culturing, filtering and magnetic separation using cell-surface antigen markers. Here we provide a comprehensive protocol on how to isolate and culture individual cell types from whole endometrial tissues for use in in vitro experiments.

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, 42 views, 0 Q&A, How to cite
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, 116 views, 0 Q&A, How to cite
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, 102 views, 0 Q&A, How to cite
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, 130 views, 0 Q&A, How to cite
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, 178 views, 0 Q&A, How to cite
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, 322 views, 0 Q&A, How to cite
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 ...

Single-cell Visualization of Chromosome Transcriptional Territories by RNA-paint

Authors: Céline Vallot
Céline VallotAffiliation 1: University Paris Diderot, Sorbonne Paris Cité, Epigenetics and Cell Fate, Paris, France
Affiliation 2: CNRS, UMR7216 Epigenetics and Cell Fate, Paris, France
For correspondence: celine.vallot@univ-paris-diderot.fr
Bio-protocol author page: a3463
 and Claire Rougeulle
Claire RougeulleAffiliation 1: University Paris Diderot, Sorbonne Paris Cité, Epigenetics and Cell Fate, Paris, France
Affiliation 2: CNRS, UMR7216 Epigenetics and Cell Fate, Paris, France
For correspondence: claire.rougeulle@univ-paris-diderot.fr
Bio-protocol author page: a3464
date: 9/5/2016, 451 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1914.

[Abstract] We developed a FISH-based method to directly assess chromosome-wide transcriptional activity, thereby enabling the visualization of the actively transcribed fraction of a chromosome at the single-cell level. We applied this method to probe the activity of X-chromosomes and its instability in the context of human embryonic stem cells and cancer cells....

Bone Marrow-derived Endothelial Progenitor Cell Intercellular Adhesion Assay

Authors: Yubo Tang
Yubo TangAffiliation 1: Department of Pharmacy, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Affiliation 2: Centre for Translational Bone, Joint and Soft Tissue Research, Medical Faculty and University Hospital Carl Gustav Carus, Technical University Dresden, Dresden, Germany
Bio-protocol author page: a3443
Angela Jacobi
Angela JacobiAffiliation: Centre for Translational Bone, Joint and Soft Tissue Research, Medical Faculty and University Hospital Carl Gustav Carus, Technical University Dresden, Dresden, Germany
Bio-protocol author page: a3444
Corina Vater
Corina VaterAffiliation: Centre for Translational Bone, Joint and Soft Tissue Research, Medical Faculty and University Hospital Carl Gustav Carus, Technical University Dresden, Dresden, Germany
Bio-protocol author page: a3445
Xuenong Zou
Xuenong ZouAffiliation: Department of Spinal Surgery, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Bio-protocol author page: a3446
 and Maik Stiehler
Maik StiehlerAffiliation 1: Centre for Translational Bone, Joint and Soft Tissue Research, Medical Faculty and University Hospital Carl Gustav Carus, Technical University Dresden, Dresden, Germany
Affiliation 2: Department of Orthopaedics, Joint and Soft Tissue Research, University Hospital Carl Gustav Carus, Dresden, Germany
For correspondence: Maik.Stiehler@uniklinikum-dresden.de
Bio-protocol author page: a3447
date: 8/20/2016, 378 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1909.

[Abstract] Cell-cell adhesion ensures tight contacts between neighbouring cells, which is necessary for cell segregation, as well as for the morphological and functional differentiation of different tissues. Evidently there are cell-cell recognition systems that make cells of the same type preferentially adherent to one another. Homotypic cell adhesion is particularly ...

Bone Marrow Mesenchymal Stem Cells Adhesion Assay

Authors: Zhigang Yang
Zhigang YangAffiliation: Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union of Medical Colleg, Beijing, China
Bio-protocol author page: a3411
 and Ran Xiao
Ran XiaoAffiliation: Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union of Medical Colleg, Beijing, China
For correspondence: xiaoran@psh.pumc.edu.cn
Bio-protocol author page: a3412
date: 8/5/2016, 392 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1895.

[Abstract] Mesenchymal stem cells (MSCs) are widespread in adult organisms and involved in tissue maintenance and repair as well as in the regulation of hematopoiesis and immunologic responses. As cell adhesion play important roles in cell interactions and signaling, to thoroughly evaluate the adhesion ability of MSCs is of vital important to clarify the mechanism ...

Neurosphere Co-culture Assay

Authors: Fanny Ehret
Fanny EhretAffiliation 1: German Center for Neurodegenerative Diseases (DZNE) Dresden, Germany
Affiliation 2: CRTD-Center for Regenerative Therapies Dresden, Genomics of Regeneration, Technische Universität Dresden, Germany
For correspondence: Fanny.Ehret@mail.de
Bio-protocol author page: a3374
Steffen Vogler
Steffen VoglerAffiliation 1: German Center for Neurodegenerative Diseases (DZNE) Dresden, Germany
Affiliation 2: CRTD-Center for Regenerative Therapies Dresden, Genomics of Regeneration, Technische Universität Dresden, Germany
Bio-protocol author page: a3375
 and Gerd Kempermann
Gerd KempermannAffiliation 1: German Center for Neurodegenerative Diseases (DZNE) Dresden, Germany
Affiliation 2: CRTD-Center for Regenerative Therapies Dresden, Genomics of Regeneration, Technische Universität Dresden, Germany
For correspondence: gerd.kempermann@crt-dresden.de
Bio-protocol author page: a3376
date: 8/5/2016, 589 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1883.

[Abstract] The hippocampal niche is one of two areas in the brain where stem cells reside. In this neurogenic niche, stem cells can be found in close proximity to astrocytes and in contact with microvessels consisting of pericytes and endothelial cells. To study the regulatory interplay of this complex niche network in a simplified in vitro model, we established ...
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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, 10307 views, 0 Q&A, How to cite
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, 8585 views, 0 Q&A, How to cite
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, 8387 views, 0 Q&A, How to cite
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, 7768 views, 0 Q&A, How to cite
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, 7597 views, 0 Q&A, How to cite
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, 7024 views, 0 Q&A, How to cite
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, 6472 views, 2 Q&A, How to cite
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, 6113 views, 3 Q&A, How to cite
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, 5956 views, 0 Q&A, How to cite
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, 5831 views, 0 Q&A, How to cite
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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