Notch Signal Pathway Expression In The Antler Stem Cells Cultured In Vitro

Antlers are the only mammalian organ that can fully regenerate, therefore, can be utilized as a model for the study of mammalian organ regeneration. Deer antlers are growing from the permanent bony protuberances of male deer, known as pedicles. Antlerogenic periosteum is the key tissue type of antler develop. It has been demonstrated that the regeneration of antlers is resulted from the proliferation and differentiation of pedicle periosteal cells, and the latter are directly differentiated from the cells of antlerogenic periosteum. Both antlerogenic and pedicle periosteal cells are called antler stem cells, and antler regeneration has been concluded as a stem cell-based process. It's an important to study antler regeneration regulation mechanism if we want to have in-depth understanding of antler stem cells. Most scholars to study proliferation and differentiation of stem cells through investigating signaling pathways. We can infer that there is one or several signaling pathways operating in AP/PP cell systems.Notch signaling pathway is an evolutionarily conserved system and plays an important role in stem cell biology including proliferation, differentiation and apoptosis mediate through binding to its'ligand, such as delta, jagged. Maintenance of stem cell proliferation and inhibition of stem cell differentiation are the major roles played by notch signaling pathway. To study these pathways in antler generation and regeneration systems, I isolated, cultured and purified deer antler stem cells including AP and PP cells, and FP cells, cell pellets as controls, pedicles dermal cells and ear cartilage cells as generic controls.Extraction of the total RNA and design the specific primers of the notch signaling pathway factors were carried out. Expression pattern of notch signaling molecules in the cultured AP, PP, FP cells and cell pellets were examined using RT-PCR. Retrieve, purification, sequence and alignment of the target fragments were conducted.: Differential expression between AP and PP cells in mRNA of notch-1, notch-2, notch-3, notch-4, dll-3, dll-4, jagged-1, jagged-2, CBF-1, hes-1 and hes-5 were detected in my study, which indicates that notch signaling pathway may be involved in the maintenance and activation of the AP and PPcells.In this thesis, I successfully obtained AP, PP, FP, pedicle dermal cells and deer ear cartilage cells. Expression pattern of notch signaling molecules in the cultured AP, PP, FP cells and cartilaginous tissue were examined at RNA level using RT-PCR. The results qualitatively reflects that notch-delta signaling pathway plays an important role in regulating proliferation and differentiation of antler stem cells, which laid the foundation forfor follow-up study.