3D organoid culture systems are increasingly employed as powerful tools for the study of human diseases. Organoids are thought to better represent the tissue-specific niche microenvironment. R-spondin-1 (RSPO1) is one of the most extensively used niche factors for culturing 3D organoids. The first published report of an organoid culture system able to promote long-term survival and multilineage differentiation of intestinal stem cells (ISCs) employed mouse Rspo1 as one of the critical factors . Since then, R-Spondin1 has been used to establish organoid cultures from the stomach, small intestine, colon, pancreas and liver from both mouse and human sources .
RSPO1 is an intestinal growth factor that works as a potent activator of the Wnt/beta-catenin signaling pathway by binding to leucine-rich repeat–containing G protein–coupled receptors (LGRs), which are expressed in tissue stem cells. RSPO1 also binds to the transmembrane E3 ubiquitin ligases RNF43 and ZNRF3 and triggers their internalization, thereby potentiating Wnt signaling.
The 293T cell line is stably transfected to express mouse R-spondin1 protein tagged with N-terminus HA epitope tag and C-terminus mouse IgG2α Fc region. R-Spondin1 expressing 293T cell line can be used to produce either purified Rspo1 or Rspo1 conditioned media. The FC region and HA tags enable ease of purification and characterization. Cell line is not the published cell line from the Kuo lab, but rather a related cell line developed in the same lab using the reference protocol.
Cell Line Description
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• Each vial contains ≥1X10⁶ viable cells.
• Cells are tested negative for HPV-16, HPV-18, Hepatitis A, B, C, Herpesvirus type 6, 7, 8 and HIV-1 & 2 viruses by PCR.
• Cells are negative for mycoplasma contamination.
• Each lot of cells is genotyped by STR analysis to verify the unique identity of the cell line.
Storage and Stability
Store in liquid nitrogen. The cells can be cultured for at least 10 passages after initial thawing without significantly affecting the cell marker expression and functionality.