Recombinant human R-spondin 1 protein (Qk006)

Summary

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R-spondin 1 (RSPO-1) is a secreted protein of the RSPO family that regulates the canonical Wnt signaling pathway [1,2]. The Wnt pathway is a highly conserved signaling pathway that is essential for various cellular processes, including cell proliferation, differentiation, and migration. This growth factor plays a crucial role in embryonic development and patterning, paticularly in the development of the gastrointestinal tract, neural tube, skin, and limbs [3]. It 1 is involved in the maintenance of stem cells and can enhance the self-renewal capacity of stem cells, promoting tissue regeneration and repair [4]. It is also involved in the sex determination and the development and maintenance of the female sex characteristics in particular the differentiation and maintenance of the ovarian tissues [5].

RSPO1 proteins are generally characterized by a core domain known as the thrombospondin type 1 (TSP1) domain, which is responsible for their signaling activity. This protein is comprised of 263 amino acids and structured as two cysteine-rich, furin-like domains [3,6]. It acts as an agonist (activator) of the Wnt signaling pathway by binding to the leucine-rich repeat-containing G-protein coupled receptors (LGR) 4-6, leading to the stabilization of β-catenin, a key component of the pathway [7].

In cell culture, this recombinant protein is used extensively for the maintenance of self-renewal and expansion of stem cells. It is also used for the culture of intestinal, hematopoietic, and epithelial stem cells [2,7,8]. Also, the culture of stem cell-derived organoids such as intestinal, pancreas liver, colon, and stomach organoids benefit from the inclusion of  RSPO1 in the media [7,9,10].

Dysregulation of the Wnt signaling pathway, including the overexpression of R-spondin 1, has been associated with tumor growth and progression in various cancers such as colon and ovarian cancer [11,12]. Research is ongoing on the role of R-spondin 1 in various biological processes and its potential as a therapeutic target for cancer treatment. Given its role in stem cell maintenance and tissue regeneration,  RSPO1 and related proteins have also been explored as potential therapeutic targets for regenerative medicine.

Technote: R-spondin 1 in pancreatic organoids

Qkine recombinant human R-spondin 1 protein replaces conditioned media in pancreatic tumor organoid culture – data from Tuveson Laboratory, Cold Spring Harbor Lab

Comparison of pancreatic organoid growth over three passages.

P-0 indicates the initial start of the culture in full growth media supplemented with Wnt3a and R-spondin 1-conditioned media.  ‘Rec. R-spondin 1’ culture conditions were created by depleting full growth media of  RSPO1-conditioned media and supplementing it with Qk006. ‘No R-spondin 1’ culture conditions were created by depleting full growth media of  RSPO1-conditioned media but not subsequently supplementing the media with an alternative source of R-spondin 1.  No differences in organoid growth have been observed when using recombinant R-spondin 1 instead of  RSPO1-conditioned media.  Experiments have been conducted by Dennis Plenker, Ph.D. in the lab of Dr David Tuveson at Cold Spring Harbor Laboratory.

Qkine R-spondin 1 treatment on urine-derived stem cells maintains cellular morphology and nuclear expression of the nephron progenitor marker, SIX2

Previous research suggests that the removal of R-spondin 1 from the culture media of urine-derived stem cells leads to loss of kidney progenitor molecules in mice. Therefore, it was hypothesized that the addition of R-spondin 1 to the culture media would result in maintenance of SIX2 progenitor molecules and thus maintenance of the undifferentiated state.

Experiments have been conducted by Lisa Nguyen (MSc) and team in the laboratory of Professor James Adjaye at the Institute for Stem Cell Research and Regenerative Medicine, University of Dusseldorf.

• What is the function of R Spondin 1?

R-spondin 1 regulates Wnt signaling and β-Catenin stabilization. Therefore, the functions of R-spondin 1 are diverse and are associated with cell proliferation and differentiation. It maintains the self-renewal capacity of stem cells and is involved in the embryonic development especially the development of the female reproductive system, gastrointestinal track, and neural tube.

• What does the Wnt pathway do?

The Wnt signaling pathway is involved in in various biological processes. It is involved in the cell fate specification, embryonic development, stem cell maintenance, tissue regeneration, synaptic function, immune response, cancer progression, and cell migration.

• What is the molecular weight of R Spondin 1?

The molecular weight is 13 kDa.

• What does the RSPO1 gene do?

The RSPO1 gene provides instructions for producing the R-spondin 1 protein.

• Where is the RSPO1 gene located?

the RSPO1 gene is located at 1q41.2.

• What is R Spondin 1 in cell culture?

It is a growth factor supplemented to cell culture media. It is used for the culture of induced pluripotent stem cells, embryonic stem cells, epithelial stem cells, and organoids such as intestinal and colon organoids.

• How big is the R Spondin protein?

The size of the R-spondin proteins, including R-spondin 1 (RSPO1), can vary slightly depending on the specific isoform and the species being considered. In humans, RSPO1 is approximatively 263 amino acids in length with a molecular weight of 13-29 kDa.