HiActi™ Cytokines Validated in Cultured Organoids

In 2021, organoids will be listed as a key project of the "14th Five-Year Plan" national key research and development plan. Organoids are so hot, and the way to obtain them has also become the focus of attention. Different organoids require different cytokines during culture. By adding cytokines that promote or inhibit specific signaling pathways, stem cells can be made to differentiate into desired organoids. The most classic cytokine culture protocol in organoid culture is WNER (respectively: Wnt-3a, Noggin, EGF and R-Spondins), and the increase or decrease combination of these 4 factors can be applied to almost all organoid culture experiments.

Cytokines commonly used in organoid culture

R-Spodin 1

Function: R-Spodin 1 belongs to the Rspo family of Wnt regulators, which includes 4 members, namely Rspo1-4. This family is a microenvironmental factor that maintains adult stem cells in multiple organs. RSPO1 is a secreted activator protein. Rspo-1 is expressed in certain regions of the developing central nervous system and is also found in the adrenal gland, ovary, testis, thyroid, and trachea.

As an activator of the Wnt signaling pathway, Rspo1 plays a role in many aspects by enhancing Wnt signaling. It is a commonly used growth medium for gastric epithelial organoids, small intestine and colon organoids, liver organoids, prostate organoids, pancreatic organoids, and breast organoids. factor.

Wnt-3a

Function: Wnt-3a belongs to the Wnt family of signaling proteins and plays an important role in maintaining the integrity of embryonic and adult tissues. The Wnt family is highly conserved and rich in cysteine. Wnts mediate the Wnt signaling pathway by binding to cell membrane receptors and/or by binding to adjacent cell membrane receptors. The Wnt signaling pathway can be divided into canonical pathway and non-canonical pathway, one is involved in the canonical signaling pathway mediated by β-catenin, including Wnt1, Wnt2, Wnt2b, Wnt3, Wnt3a, Wnt7a, Wnt8, Wnt8b and Wnt10a; the other is not Noncanonical signaling pathways dependent on β-catenin, including Wnt4, Wnt5a, Wnt6 and Wnt11. As the most representative signaling protein of the Wnt family, Wnt3a is widely distributed and plays a key role in regulating pleiotropic cellular functions, including self-renewal, proliferation, differentiation and motility.

EGF

Function: EGF is a potent growth factor that stimulates multiple epithelial development, angiogenesis, inhibition of gastric acid secretion, fibroblast proliferation, and colony formation of epidermal cells in culture. EGF is a founding member of the EGF family, which also includes TGF-α, two regulatory proteins (AR), β-cell protein (BTC), epidermal protein (EPR), heparin-binding EGF-like growth factor (HB-EGF), Epiproteins and neuregulins (NRG1-6). Physiologically, EGF exists in various body fluids, including blood, milk, urine, saliva, semen, pancreatic juice, cerebrospinal fluid and amniotic fluid. EGF is a high-affinity ligand for the EGF receptor (ErbB). EGF binds and induces the dimerization of the EGF receptor, leading to the activation of the protein tyrosine kinase signaling pathway, thereby transducing the EGF family signal.

Noggin

Function: Noggin can inhibit TGF-β family ligands and prevent them from binding to corresponding receptors. Noggin was originally discovered as an antagonist of BMP-4 and was then shown to modulate the activity of other BMPs (BMP-2, 7, 13 and 14). Noggin is highly conserved among vertebrates. Secreted Noggin proteins regulate the activity of bone morphogenetic proteins during development. Role of the BMP signaling antagonist Noggin in the development of osteolytic bone metastases from prostate cancer.

 bFGF/FGF-2

Function: bFGF, a member of the FGF superfamily of mitogenic proteins. It is a highly specific chemotactic and mitogenic factor for many cell types and appears to be involved in the remodeling of damaged tissues such as ulcer healing, vascular repair, traumatic brain injury. bFGF is an important component of human embryonic stem cell culture medium. Furthermore, the bFGF protein is a heparin-binding cationic protein involved in various pathological conditions, including angiogenesis and the growth of solid tumors. Therefore, bFGF is considered a target for cancer chemoprevention and therapeutic strategies. bFGF is often used as an important component of cell culture medium, such as human embryonic stem cell culture medium, serum-free culture system.

FGF-10

Function: Fibroblast growth factor 10 (FGF-10) is a heparin-binding growth factor that belongs to the FGF family. Proteins of this family play central roles in prenatal development, postnatal growth and regeneration of various tissues by promoting cell proliferation and differentiation. FGF-10 is secreted by mesenchymal cells and binds to extracellular FGF-BP, FGF-10 preferentially binds and activates epithelial FGF R2 (IIIb), and interacts weakly with FGF R1 (IIIb).

 
Quality management system of HiActi™ cytokine

Quality Management System

• Certified by ISO 13485:2016, cytokines are manufactured through a fully enclosed ultra-clean production process to ensure the cleanliness and stability of proteins.
• Comply with the system of "Good Manufacturing Practice for Drugs"
• Have a Certificate of Approval Quality (COA)
• Aseptic technology and secondary sterilization filtration
• Pharmaceutical product raw material production base

Quality Assurance Management

• Special person is responsible for the management personnel training system
• Establish equipment maintenance and calibration procedures
• Raw material and supplier management system
• Complete batch production records, batch inspection records, packaging records, etc.
• Quality document records have been audited and inspected by QA
• Complete batch production and inspection records
• Validation of analytical methods

Quality Control System

• Comply with the third general rule of the 2020 edition of the Pharmacopoeia of the People's Republic of China: quality control of raw materials and excipients for the production of biological product

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.

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