A protocol to quantify mammary early common progenitors from long-term mammosphere culture

Autor: Véronique Maguer-Satta, Emmanuel Delay, Elodie Bachelard-Cascales, Marion Chapellier
Přispěvatelé: Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Cancérologie de Lyon ( CRCL ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS )
Jazyk: angličtina
Rok vydání: 2012
Předmět:
MESH: Neoplasm Proteins
Time Factors
Cellular differentiation
Cell Culture Techniques
MESH : Aged
MESH : Receptors
Cell Surface
MESH : Blotting
Western
MESH : Interleukin-3 Receptor alpha Subunit
MESH : Antineoplastic Agents
Phytogenic
[ SDV.CAN ] Life Sciences [q-bio]/Cancer
MESH: Hematopoietic Stem Cells
Mice
0302 clinical medicine
MESH: Aged
80 and over
MESH : Indoles
MESH : Tumor Cells
Cultured
MESH : Female
MESH : GTP-Binding Proteins
MESH: Glycogen Synthase Kinase 3
MESH: Maleimides
MESH: Receptors
Cell Surface
MESH: Etoposide
MESH : Maleimides
MESH: Aged
MESH: Indoles
MESH : Cell Survival
0303 health sciences
MESH: Middle Aged
Stem Cells
MESH: Protein Phosphatase 2
Cell Differentiation
General Medicine
MESH : Adult
Cell biology
MESH: Cell Survival
MESH: Young Adult
030220 oncology & carcinogenesis
MESH : Antigens
CD34
Stem cell
MESH: Cells
Cultured
Adult
MESH: GTP-Binding Proteins
MESH : Protein Phosphatase 2
MESH : Glycogen Synthase Kinase 3
MESH : Male
MESH : Sex Factors
MESH: RNA Interference
MESH: Antineoplastic Agents
Phytogenic
MESH : Young Adult
[SDV.CAN]Life Sciences [q-bio]/Cancer
MESH : Etoposide
Biology
MESH: Cell Adhesion
Colony-Forming Units Assay
03 medical and health sciences
MESH : Neoplasm Proteins
MESH : Neoplastic Stem Cells
MESH: Sex Factors
Spheroids
Cellular
MESH : Cells
Cultured
MESH: Leukemia
Animals
Humans
MESH: Blotting
Western
MESH : Middle Aged
MESH: Tumor Cells
Cultured
Progenitor cell
Mammary Glands
Human
MESH : Aged
80 and over
030304 developmental biology
Progenitor
MESH: Interleukin-3 Receptor alpha Subunit
MESH: Humans
MESH : Hematopoietic Stem Cells
Stem cell population
MESH : Humans
MESH : Leukemia
MESH: Adult
Cell Biology
MESH: Antigens
CD34
MESH: Neoplastic Stem Cells
MESH: Male
MESH : Cell Adhesion
Cell culture
Immunology
NIH 3T3 Cells
MESH : RNA Interference
MESH: Female
Developmental Biology
Zdroj: Oncogene
Oncogene, Nature Publishing Group, 2012, Chapter 1 (6), pp.Unit 1E.7. ⟨10.1002/9780470151808.sc01e07s20⟩
Current Protocols in Stem Cell Biology
Oncogene, Nature Publishing Group, 2012, Chapter 1 (6), pp.Unit 1E.7. 〈10.1002/9780470151808.sc01e07s20〉
ISSN: 0950-9232
1476-5594
DOI: 10.1002/9780470151808.sc01e07s20⟩
Popis: International audience; Here we describe a protocol established in our laboratory to quantify early common progenitors/stem cells grown as spheres in non-adherent culture conditions. This protocol is based on the combination of two functional tests: the mammosphere assay to maintain and enrich early mammary progenitors/stem cells, and the epithelial colony-forming cells (E-CFC) assay to identify and quantify further differentiated epithelial progenitors. Primary spheres mainly contain progenitors and rare stem/early common progenitor cells while secondary and tertiary spheres contain progenitor cells derived from the early common progenitor/stem cell population maintained through passages and partially differentiated. Spheres are enzymatically and mechanically dissociated; the derived cells are subsequently plated on irradiated NIH-3T3 fibroblasts for further processing, as in the E-CFC assay. The principle of this assay is to quantify the number of epithelial colonies generated by cells present in the different sequential spheres. This assay has therefore been named ECP-DC, standing for Early Common Progenitor-Derived Colonies assay.
Databáze: OpenAIRE
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