iPSC-hDPSC

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.

37°C, 5% CO₂, humidified atmosphere.

To improve the cell detachment and reseeding process, follow these steps:

1. Remove the spent medium and rinse the cells with PBS (without calcium and magnesium).
2. Add the detaching solution and allow it to act as per the manufacturer's instructions.
3. Carefully add fresh medium to the cells.
4. Gently dissociate the cells to ensure minimal damage.
5. Coat a fresh 6-well culture plate with vitronectin and seed the dissociated cells into two wells for further cultivation.

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.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.