Cultrex® R-spondin1 (Rspo1) Cells

The 293T cell line is stably transfected to express murine Rspo1 with an N-terminal HA epitope tag and fused to a C-terminal murine IgG2a Fc fragment. This cell line is used to produce either purified Rspo1 or Rspo1 conditioned media. The murine Rspo1 protein has been used extensively in organoid culture to maintain Lgr5+ stem cells, and the Fc and HA tags make it easy to purify or characterize.

Available Size(s): 1 Vial (0.5 ml), 1x10^6 cells Catalog Number: 3710-001-K Categories: ,

Description

NOTE: Starting January 1st, 2017 free sample sizes of Cultrex® Organoid Harvesting SolutionCultrex® BME Type 2,  and Cultrex® BME Type R1  will be added to your order.  These products are also sold individually, more information on Trevigen®’s Cultrex Organoid products can be found here.

Roof plate-specific Spondin-1 (R-Spondin 1 or Rspo1), also known as CRISTIN3, is a 27 kDa secreted activator protein that belongs to the R-Spondin family. R-Spondins positively regulate Wnt/beta-catenin signaling, most likely by acting as a ligand for LGR4-6 receptors and an inhibitor for ZNRF3. R-Spondin-1 induces proliferation of intestinal crypt epithelial cells, increases intestinal epithelial healing, and supports intestinal epithelial stem cell renewal.

Features:

  • Robust and stable expression of R-Spondin1.
  • Two epitope tags provide flexibility to purify and detect the expressed protein.
  • Accesible source of either purified R-Spondin1 or R-Spondin1 conditioned medium.
  • Suitable to supplement both human and murine organoid culture medium.

Quality control specifications:

  • Negative by PCR for Mycoplasma.
  • Expression detected by Western Blot – A band of approximately 70 kDa corresponding to HA-R-Spondin1-Fc is detected in the conditioned medium of cells after 7 days of culture.
  • Biological activity determined for each lot:
    • TopFlash assay – HA-R-Spondin1-Fc must induce the expression of luciferase under the control of a TCF/LEF promoter.

Protocol(s)

TopFlash Assay

protocol_3710-001-01

protocol_3710-001-K

Material Safety Data Sheet(s)

msds_3710-001-01 HA-R-Spondin1 293T Fc Cell line

Technical Tips(s)

Harvesting organoids for biochemical analysis

Immunofluorescence of organoids embedded in Basement Membrane Matrix

Production of R-Spondin 1 conditioned medium using Cultrex® Rspo1 cells

Production of R-Spondin1 for organoid culture.  The 293T cell line is stably transfected to express murine Rspo1 with an N-terminal HA epitope tag and fused to a C-terminal murine IgG2a Fc fragment. A) The HA-R-Spondin1-Fc 293T cell line is cultured with Zeocin to select for stably transfected cells. B) Production of HA-R-Spondin1-Fc is characterized using Western Blot for R-Spondin1 protein (arrow). C) HA-R-Spondin1-Fc induces activation of Wnt/ß-catenin response when evaluated using the TopFlash Luciferase assay.