干细胞实验指南
Stem Cell Protocols
Rat Cortical Stem Cell Culture Expansion Protocol
This protocol must be read in its entirety before using Catalog # NSC001. Caution: The rat cortical stem cells used in this protocol contain trace amounts of human transferrin and DMSO, and the media used in this protocol contains trace amounts of transferrin. The transferrin was tested at the donor level using an FDA licensed method and found to be non-reactive for anti HIV-1/2 and Hepatitis B surface antigen.
Mouse Hematopoietic Lineage Depletion
Antibodies including anti-mouse CD5, CD11b, CD45R (B220), Ly-6G (Gr-1), and TER-119 can be used to efficiently tag bone marrow-derived cells committed to major hematopoietic lineages, including T lymphocytes, B lymphocytes, monocytes/macrophages, granulocytes, and erythrocytes. These antibodies can be used in conjunction with magnetic particle separation systems or flow cytometric cell sorting to deplete lineage-committed cells and enrich for uncommitted hematopoietic progenitors.
Culturing Rat Cortical Stem Cells: Expansion using the Monolayer System
Ex vivo expanded neural stem cells serve as excellent tools for researchers studying neural development and neurological disorders. Ready-to-use primary cortical stem cells, isolated from E14.5 Sprague-Dawley rats (Catalog # NSC001), can be grown in monolayer, as described here, or as neurospheres. These cells retain capacity for multi-lineage differentiation into astrocytes, neurons, and oligodendrocytes.
Please read the protocol in its entirety before starting.
Definitive Endoderm Differentiation of BG01V Human Embryonic Stem Cells
This protocol is designed for use with BG01V human embryonic stem cells grown in Mouse Embryonic Fibroblast (MEF) Conditioned Media (AR005). If using different cell lines or growth media, the protocol below may need to be modified. The quality of the human pluripotent cells used in the differentiation is critical. Use of suboptimal quality or very high passage pluripotent cells can result in decreased differentiation efficiency and/or increased cell death.
Endoderm, Ectoderm, and Mesoderm Differentiation of BG01V Human Embryonic Stem Cells
This protocol is designed for BG01V human embryonic stem (hES) cells grown in Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005). If using different cell lines or growth media, the protocol below may need to be modified.
The quality of the human pluripotent cells used in the differentiation is imperative. Use of suboptimal quality or very high passage pluripotent cells can result in decreased differentiation efficiency and/or increased cell death.
Chondrogenic Differentiation of Human/Mouse Mesenchymal Stem Cells
Mesenchymal stem cells (MSCs) are functionally defined by their capacity to self renew and their ability to differentiate into multiple cell types including adipocytes, chondrocytes and osteocytes. This protocol describes the chondrogenic differentiation of MSCs using the StemXVivo Human/Mouse Chondrogenic Base Media (Catalog # CCM005) and StemXVivo Human/Mouse Chondrogenic Supplement (Catalog # CCM006).
The Human Colony Forming Cell (CFC) Assay using Methylcellulose-based Media
The colony forming cell (CFC) assay, also referred to as the methylcellulose assay, is an in vitro assay used in the study of hematopoietic stem cells. The assay is based on the ability of hematopoietic progenitors to proliferate and differentiate into colonies in a semi-solid media in response to cytokine stimulation. The colonies formed can be enumerated and characterized according to their unique morphology.
Please read the protocol in its entirety before starting.
Procedure for Serum-Free Expansion of Human Mesenchymal Stem Cells
Mesenchymal stem cells (MSCs) are functionally defined by their capacity to self renew and their ability to differentiate into multiple cell types including adipocytes, chondrocytes, and osteocytes (1, 2). MSCs are phenotypically characterized as CD105+, CD90+, CD73+, CD44+, CD34-, CD45-, and CD11b- cells. This protocol describes the expansion of human MSCs using the StemXVivo™ Serum-Free Human MSC Expansion Media (R&D Systems, Catalog # CCM014).
Adipogenic Differentiation of Human/Mouse Mesenchymal Stem Cells
Mesenchymal stem cells (MSCs) are functionally defined by their capacity to self renew and their ability to differentiate into multiple cell types including adipocytes, chondrocytes, and osteocytes. This protocol describes the adipogenic differentiation of human and mouse MSCs using the StemXVivo™ Human/Mouse Osteogenic/Adipogenic Base Media (Catalog # CCM007) and StemXVivo Adipogenic Supplement (Catalog # CCM011).
Please read the protocol in its entirety before starting.
Procedure for the Expansion of Human Mesenchymal Stem Cells
Mesenchymal stem cells (MSCs) are functionally defined by their capacity to self renew and their ability to differentiate into multiple cell types including adipocytes, chondrocytes, and osteocytes (1, 2). MSCs are phenotypically characterized as CD34-, CD105+, and CD90+ cells. This protocol describes the expansion of human MSCs using the StemXVivo™ MSC Expansion Media (R&D Systems, Catalog # CCM004).
Please read the protocol in its entirety before starting.