iPSC-hDPSC

Categories: iPSC Description: The iPSC-hDPSC cell line represents a cutting-edge model derived from human dental pulp stem cells (hDPSCs). These cells originate from dental pulp tissue, where they undergo enzymatic digestion to isolate the adherent fraction. Upon reaching 70-80% confluence, the cells are subcultured and cryopreserved at Passage 1. This foundational step is crucial in maintaining the cellular integrity and ensuring the viability of the stem cells for future applications. The reprogramming of hDPSCs into induced pluripotent stem cells (iPSCs) is a significant advancement, achieved through an episomal reprogramming technique. This method employs vectors that incorporate the Yamanaka factors-Oct-4, Sox-2, and Klf-4-alongside p53 Anti-sense, EBNA-1, and a Red Fluorescent Protein marker. The episomal approach is particularly advantageous as it avoids the integration of reprogramming factors into the genome, thus preserving the genetic stability of the iPSCs. The resultant iPSC-hDPSC line exhibits pluripotent characteristics, making it a valuable tool for research in regenerative medicine, disease modeling, and drug discovery. However, it is important to note that this cell line is intended solely for research purposes and is not suitable for therapeutic or in vivo applications. Organism: Human Tissue: Third molar Disease: Normal donor Ethnicity: Caucasian Morphology: iPSC, colonies with defined edges Cell Type: Stem cells Growth Properties: Adherent Citation: iPSC-hDPSC (Cytion catalog number 300622) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_B7DW Culture Medium: mTESR Dissociation Reagent: ReLeSR Subculturing: To improve the cell detachment and reseeding process, follow these steps: Remove the spent medium and rinse the cells with PBS (without calcium and magnesium). Add the detaching solution and allow it to act as per the manufacturer's instructions. Carefully add fresh medium to the cells. Gently dissociate the cells to ensure minimal damage. Coat a fresh 6-well culture plate with vitronectin and seed the dissociated cells into two wells for further cultivation. Seeding Density: To ensure optimal cell growth and experimental consistency, it is recommended to use a seeding density of 5,000 to 10,000 cells/cm2 once the cells have adapted to the culture conditions. Fluid Renewal: Daily Post Thaw Recovery: Fast Freeze Medium: As a cryopreservation medium, use 80% FBS + 10% basal medium + 10% DMSO to maintain viability, or CM-1 (Cytion catalog number 800100) for superior cryoprotection, preventing unwanted differentiation while preserving pluripotency and cellular integrity. Thawing And Culturing Cells: Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks, for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes. Sterility: Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 3: 860015.0 Required Product 4: 830100.0