Neuronal Cells Directed Differentiation Technology
Creative Biolabs combines many years of research experience in nerve and stem cell (SC) fields to provide customers with a one-stop service for targeted differentiation of neural cells. SCs, also known as "omnipotent cells", are characterized by rapid proliferation, multidirectional differentiation, low immunity, and self-repair. Cells directionally differentiated from SCs have the characteristic properties of neural stem cells (NSCs), which can differentiate into neurons and glia. These nerve cell types play an important role in the study of human nervous system development and disease.
The difficulty in regeneration and recovery of the central nervous system after nerve injury has always been a hotspot and difficulty in neuroscience and regenerative medicine. NSCs come from a limited number of sources, but SCs come from a wide range of sources. Inducing SCs to differentiate into nerve cells is of great significance for the development of cell therapy. At present, SCs transplantation is widely used in regenerative medicine treatment of neurological diseases. SCs have the potential of self-renewal and multidirectional differentiation, whose differentiation direction is affected by many factors such as genes, cytokines, and the internal environment. SCs can be induced to differentiate into nerve cells and are used to establish disease-related cell types and repair nervous system dysfunction. However, studies have found that SCs transplantation in vivo induces immune rejection and misdifferentiated safety issues. Therefore, in vitro induction of SCs directed neural differentiation is of great significance for the treatment of neurological diseases. Among them, induced pluripotent stem cells (iPSCs) completely overcome the ethical and immune rejection problems of embryonic stem cells (ESCs) in clinical application, have important clinical application value, and become the main cell source of SCs therapy.
Inducing SCs to Differentiate into NSCs
NSCs play an important role in neural development and repair of damaged nerve tissue. In addition, NSCs have certain practical value in judging the efficacy and toxicity of drugs. Creative Biolabs provides a cell culture medium containing specific inducers to induce the differentiation of SCs into NSCs, and the expression of neurospecific markers Nestin and SRY-Box Transcription Factor 2 (SOX2) can be verified by immunofluorescence staining. Once NSCs differentiate into neurons and glial cells, Nestin expression stops.
Fig.1. Cell culture and induction of NSCs from iPSCs. (Yuan, 2013)
Fig.2. The expression of pluripotent potent and neural markers during the induction process of the iPSCs to NSCs. (Yuan, 2013)
Inducing NSCs to Differentiate into Neurons
Cytokines play an important role in the differentiation of NSCs. Neurotrophic factor affects the differentiation of NSCs into terminal cells. If cultured NSCs are exposed to brain-derived neurotrophic factors, large numbers of NSCs can exhibit the properties of differentiated neurons. Neuronal cells are the main undertaker of nervous system functional activities and have the function of feeling stimulation and conducting excitement. Creative Biolabs can differentiate enough neurons for subsequent detection. Once NSCs differentiate into neuron cells, neuron cells specifically express microtubule associated protein 2 (Map2) and neuronal class III beta-tubulin (Tuj-1).
Fig.3. Immunofluorescence staining of Map2 and Tuj-1 (neuronal markers) in mature neurons. (Ebadi, 2018)
Inducing NSCs to Differentiate into Astrocytes
Glial cells perform auxiliary functions such as supporting, protecting, and nourishing neurons. Astrocytes play important roles in development, blood flow regulation, synaptic transmission, and energy metabolism. Loss or abnormality of astrocyte function is associated with a variety of neurodegenerative disease processes. Creative Biolabs can provide a one-stop solution to our customers' astrocyte acquisition and purification problems. The most specific markers of astrocytes are glial fibrillary acidic protein (GFAP) and S100 calcium-binding protein B (S100β).
Fig.4. Immunostaining of FACS purified astrocytes with astrocyte markers S100β, GFAP and reporter gene GFP. (Zhang, 2016)
Creative Biolabs Services
|SCs are differentiated into NSCs||The expression of Nestin and SOX2 is detected by immunofluorescence staining||6 weeks|
|NSCs are differentiated neurons||The expression of Map2 and Tuj-1 is detected by immunofluorescence staining||3-6 weeks|
|NSCs are differentiated astrocytes||The expression of GFAP and S100β is detected by immunofluorescence staining||3-6 weeks|
Creative Biolabs can provide customers with efficient technology services on SCs directed differentiation to nerve cells. Please contact us to discuss your project. Our professional team has expertise and experience in the induction and differentiation of SCs. We rely on a mature SCs technology platform to carry out personalized targeted differentiation research of nerve cells for you.
- Yuan, T.; et al. Human induced pluripotent stem cell-derived neural stem cells survive, migrate, differentiate, and improve neurologic function in a rat model of middle cerebral artery occlusion. Stem Cell Research & Therapy. 2013, 4(3):73.
- Ebadi, R.; et al. Comparison of two different media for maturation rate of neural progenitor cells to neuronal and glial cells emphasizing on expression of neurotrophins and their respective receptors. Molecular Biology Reports. 2018, 45(6):2377-2391.
- Zhang, PW.; et al. Generation of GFAP: GFP astrocyte reporter lines from human adult fibroblast-derived iPS cells using zinc-finger nuclease technology. Glia. 2016, 64(1):63-75.