Human Wnt3a expressed in HEK293 cells exhibites high activity promoting organoids cultivation

Organoids are in vitro-created three-dimensional models closely resembling natural tissues or organs, and they hold significant significance within the realms of disease modeling, organ rejuvenation, and individualized medical care. The cultivation of these organoids necessitates the careful management of a diverse array of cytokines, with Wnt3a standing out as a frequently employed factor that assumes a pivotal role in nurturing organoids derived from a wide range of tissues.

What is Wnt-3a? 

Wnt-3a belongs to the Wnt family, a group of secreted proteins comprising 19 human variants, including Wnt1, Wnt2, Wnt2b (Wnt13), Wnt3, Wnt3a, Wnt4, Wnt5a, Wnt5b, Wnt6, Wnt7a, Wnt7b, Wnt8a, Wnt8b, Wnt9a (also known as Wnt14), Wnt9b (Wnt14b), Wnt10a, Wnt10b, Wnt11, and Wnt16. This family is characterized by its conservation and cysteine-rich composition. Wnt proteins facilitate the Wnt signaling pathway by binding to cell membrane receptors or neighboring cell membrane receptors. This pathway can be categorized into the canonical pathway, mediated by β-catenin, which includes Wnt1, Wnt2, Wnt2b, Wnt3, Wnt3a, Wnt7a, Wnt8, Wnt8b, and Wnt10a, and the non-canonical pathway, independent of β-catenin, which encompasses Wnt4, Wnt5a, Wnt6, and Wnt11. Wnt3a, as the quintessential member of the Wnt family, is ubiquitously distributed and exerts a pivotal role in governing various cellular functions, such as self-renewal, proliferation, differentiation, and motility.

Wnt-3a Signaling Pathway Transduction Mechanism 

The Wnt signaling pathway is a multifaceted network of protein interactions primarily associated with embryonic development and cancer. However, it also plays a role in normal physiological processes in adult organisms. Wnt3a binds to its receptor, the Frizzled (Fz) protein, along with the co-receptor lipoprotein receptor-related protein (LRP5/6), subsequently activating the scaffolding protein Disheveled (Dvl). This activation leads to the disassembly of the "destruction complex," which consists of Casein kinase 1 (CK1), glycogen synthase kinase-3β (GSK-3β), and scaffolding proteins adenomatous polyposis coli (APC), Axin1, and Axin2. Dissolving this complex stabilizes the free β-Catenin protein in the cytoplasm. The accumulated β-Catenin in the cytoplasm then translocates into the nucleus, where it binds to transcription factors of the TCF/LEF family, ultimately activating downstream target genes such as c-myc and Cyclin D1. This cascade induces subsequent cellular responses. In summary, the Wnt signaling pathway can be simplified as follows: Wnt3a→FZD→DVL→complex dissolution→β-Catenin enters the nucleus→TCF/LEF→downstream gene transcription. Without Wnt3a, β-catenin is normally degraded in the cytoplasm by the "destruction complex," which targets it for ubiquitination and subsequent degradation in the proteasome. As a result, it cannot enter the nucleus to initiate the transcription of downstream genes. The Wnt/β-catenin signaling pathway is highly conserved throughout evolution and plays a fundamental role in embryonic development, cell growth, differentiation, polarity formation, neural development, and carcinogenesis.

 Wnt3a and Organoids 

Wnt3a holds a central position as one of the most frequently employed culture factors for organoid generation. Its high activity, consistency between batches, lack of contamination, and serum-free preparation are key factors contributing to the success of organoid experiments. Wnt3a has found widespread use in organoid cultures of various tissues, including the intestine, stomach, pancreas, liver, thyroid, esophagus, and nervous system. In an effort to assist scientific researchers and pharmaceutical companies in their studies on the clinical application of Wnt3a in organoid formation and tumor research, YEASEN has introduced a highly active Wnt3a protein product (Catalog: 92276ES) designed to facilitate organoid culture.

Figure 2. Human wnt3a Protein on SDS PAGE under reduced condition. 

Figure 3. Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells. The ED50 for this effect is 25 ng/mL.

Organoid Culture Validation

Yeasen cytokines have been verified on human tumor organoids such as lung cancer, cervical cancer, ovarian cancer, pancreatic cancer and intestinal cancer, and normal organoids such as human and mouse small intestine organoids.

Related Cytokines information