General description
OP9-DL4-7FS is a stable bone marrow derived stromal cell line expressing the Notch ligand- Delta like 4 (DL4) and human IL-7, FLT3L, and SCF (7FS) ectopically.
Notch signaling controls multiple cell-fate decisions. Four Notch receptors (Notch 1- Notch 4) have been identified in mammals. These receptors interact with Jagged or Delta-like (DL) family members of Notch ligands. This is followed by cleavage of the intracellular domain of Notch and its subsequent translocation into the nucleus, where it binds with transcription factors and activates transcription of various downstream target genes. Notch signaling is essential in early T- cell lineage commitments. Bone marrow progenitor cells expressing constitutively-active Notch develop into CD4 and CD8 double positive T cells rather than B cells. They also play an important role in development of CD4+ and CD8+ single positive cells from double positive precursor thymocytes, in addition to development of T-cells with αα+ TCRs. Of the 4 Notch receptors, Notch1 (N1) receptor signaling has been shown to be sufficient for T cell development. The delta like ligands are physiologically relevant N1 ligands. DL1 interacts with both N1 and Notch 2 (N2) to induce T- cell lineage commitment. However, DL4 interacts specifically only with N1 and supports T cell commitment and maturation both in vitro and in vivo. Moreover, results from previous binding studies show that binding between DL1 and N1 is weak as opposed to the stronger binding between DL4 and N1. Of the DL1 and DL4 delta like ligands, DL4 exhibits greater capacity to activate the Notch pathway in hematopoietic progenitor cells.
Human hematopoietic stem cells (HSCs) require additional cytokines for survival and propagation. Three important components are added separately to HSC/OP9-DL cocultures. These components are Interleukin-7 (IL-7), stem cell factor (SCF), and FMS-like tyrosine kinase 3 ligand (FLT3L). To circumvent the costly addition of these cytokines, an improved OP9-DL4 cell line was designed and designated OP9-DL4-7FS.5 This improved OP9 cell line was transduced to express human IL-7, FLT3L, and SCF (7FS), making it a highly cost-efficient cell line and improved version of the original OP9-DL4 cell line. In addition, the cerulean fluorescent protein (CeFP) gene was substituted for GFP using restriction enzymes to distinguish the new “7FS” cell line from the original OP9-DL4 cell line which had been sorted for GFP expression.
Source
OP9-DL4-7FS cells were genetically modified from OP9 stromal cells derived from mouse bone marrow.
References
1. Methods Mol Biol 2023, 2580:249-260.
2. Immunity 2002, 17(6): 749-756.
3. J Orthop Sci. 2005, 10(6): 589-594.
4. Curr Opin Immunol. 2007, 19(2): 163-168.
5. Cold Spring Harb Protoc. 2009, 2009(2): pdb.prot5156.
Application
Each vial contains > 1X106 viable cells.
Cells are tested negative for infectious diseases by a Mouse Essential CLEAR Panel by Charles River Animal Diagnostic Services.
Cells are verified to be of mouse origin and negative for interspecies contamination from human, rat, Chinese hamster, Golden Syrian hamster, and nonhuman primate (NHP) as assessed by a Contamination Clear panel by Charles River Animal Diagnostic Services
Cells are negative for mycoplasma contamination.
Features and Benefits
OP9-DL4-7FS is a stable bone marrow derived stromal cell line expressing the Notch ligand-Delta like 4 (DL4) and human IL-7, FLT3L, and SCF (7FS) for stem cell differentiation applications.
Preparation Note
OP9-DL4-7FS cells should be stored in liquid nitrogen. The cells can be cultured for at least 10 passages without significantly affecting cell marker expression and function.
Other Notes
Subject to local law, this product is intended to be sold for internal in vitro research use only subject to terms and conditions found here: www.sigmaaldrich.com/restrictedcelluse. This product may not be: re-engineered or copied; used to make derivatives, modifications or functional equivalents; used to obtain patents or other IP claiming use of the product; used to develop, test, or manufacturer a commercial product; used as a component in a commercial product; resold or licensed; used in any clinical applications or trials; or used in humans. A license or limited commercial use agreement is required for use by any for-profit entity, use in services, and use in sponsored academic research. For information regarding any such use, please contact
[email protected].
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
