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Questions and Answers
What is the most widely used method of cell preservation?
What is the most widely used method of cell preservation?
Hypothermic storage does not affect cell function.
Hypothermic storage does not affect cell function.
False
What component is tested in specialized media for hypothermia?
What component is tested in specialized media for hypothermia?
Iron chelators
Cryopreservation is the most widely used method of cell preservation, while _______ offers an alternative long-term storage solution.
Cryopreservation is the most widely used method of cell preservation, while _______ offers an alternative long-term storage solution.
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Match the preservation method with its description:
Match the preservation method with its description:
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What is the primary challenge associated with transport for cell preservation?
What is the primary challenge associated with transport for cell preservation?
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Processing cells on-site can include thawing and washing.
Processing cells on-site can include thawing and washing.
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What is one of the key characteristics that an ideal preservation method should maintain?
What is one of the key characteristics that an ideal preservation method should maintain?
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The challenge of __________ involves managing delays, vibrations, and temperature fluctuations during transport.
The challenge of __________ involves managing delays, vibrations, and temperature fluctuations during transport.
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Match the following preservation techniques with their appropriate duration:
Match the following preservation techniques with their appropriate duration:
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Which of the following is NOT a level of investment required in a clinical setting for cell therapies?
Which of the following is NOT a level of investment required in a clinical setting for cell therapies?
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Cell recovery after preservation requires extensive manipulation of cells in the clinic.
Cell recovery after preservation requires extensive manipulation of cells in the clinic.
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Name one type of specialized clinic that may focus on cell therapies.
Name one type of specialized clinic that may focus on cell therapies.
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What is the shelf life of refrigerated RBCs?
What is the shelf life of refrigerated RBCs?
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Fresh frozen plasma (FFP) must be used within 48 hours of thawing and storing at 4ºC.
Fresh frozen plasma (FFP) must be used within 48 hours of thawing and storing at 4ºC.
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What challenges are associated with the storage of blood products?
What challenges are associated with the storage of blood products?
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Platelets have a shelf life of ___ days from donation.
Platelets have a shelf life of ___ days from donation.
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Match the blood products with their respective storage conditions:
Match the blood products with their respective storage conditions:
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What is one factor that determines the transportation of cell products?
What is one factor that determines the transportation of cell products?
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Allogeneic cell therapy involves using cells from the same individual receiving the therapy.
Allogeneic cell therapy involves using cells from the same individual receiving the therapy.
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What is a significant challenge when it comes to storing blood products?
What is a significant challenge when it comes to storing blood products?
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Match the cell preservation strategies with their descriptions:
Match the cell preservation strategies with their descriptions:
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Match the transportation challenges with their descriptions:
Match the transportation challenges with their descriptions:
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Match the processes involved in cell therapy with their functions:
Match the processes involved in cell therapy with their functions:
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Match the novel harvesting methods with their characteristics:
Match the novel harvesting methods with their characteristics:
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Match the cell detachment techniques with their processing steps:
Match the cell detachment techniques with their processing steps:
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Match the cell preservation methods with their descriptions:
Match the cell preservation methods with their descriptions:
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Match the challenges associated with the transportation of cell products:
Match the challenges associated with the transportation of cell products:
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Match the components of cell therapy processes with their descriptions:
Match the components of cell therapy processes with their descriptions:
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Match the novel harvesting methods with their specific techniques:
Match the novel harvesting methods with their specific techniques:
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Match the cell detachment techniques with their purposes:
Match the cell detachment techniques with their purposes:
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Study Notes
Transport in Cell Therapy
- Banking benefits include consistent starting material and no need for continuous culture, reducing costs and risks of phenotypic/genotypic changes and contamination.
- Example blood products include red blood cells (RBCs), fresh frozen plasma (FFP), and platelets, each with specific handling and storage requirements.
Blood Product Handling
-
RBCs:
- Refrigerated with a shelf life of up to 42 days.
- Stock balance in hospitals is crucial to minimize waste, with infusion required within 30 minutes of fridge removal.
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Fresh Frozen Plasma (FFP):
- Frozen, to be defrosted onsite.
- Must be used within 24 hours if stored at 4ºC post-thaw.
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Platelets:
- Shelf life is approximately 5 days from donation (often closer to 3 days).
- Stored at blood banks and require a hospital request for use.
Decision Factors in Cell Therapy
- Cell therapy can be autologous or allogeneic; the scale of production further complicates decision-making.
- Business models can involve single or multi-site manufacturing and on or off-site processes.
- The preservation method directly influences transportation logistics, system flexibility, and at-clinic processing capabilities.
Storage Challenges
- Storage systems can be integrated into manufacturing sites, but short shelf life impacts quality control and business models.
- Issues can arise from contamination (e.g., leaky bags/vials, liquid nitrogen) and understanding how storage affects products.
- Backup systems are essential for maintaining product integrity.
Transportation Challenges
- Common transportation methods include refrigerated and frozen transport utilizing dry ice or dry shippers, although the latter can be costly and labor-intensive.
- Critical considerations involve understanding how transportation delays, vibrations, and temperature fluctuations affect product viability.
Clinic Processing Considerations
- Determine feasibility of on-site processing, such as thawing and washing, and its effects on cell viability and function.
- Matching the availability of cells, surgeons, and patients is vital for successful outcomes.
- Level of investment in equipment and staff training is necessary for effective cell therapy implementation.
Preservation Methods
- Various preservation strategies include cryopreservation, desiccation, and hypothermia, each with specific applications and limitations.
- Cryopreservation is favored for its widespread applicability and compatibility with Good Manufacturing Practice (GMP).
- An ideal preservation method should maintain biological characteristics, support transportation stability, and require minimal manipulation during reactivation.
Hypothermic Storage Insights
- Specialized media during hypothermic storage may yield variable recovery outcomes depending on medium composition.
- Research shows issues with cell function retention, with studies indicating only partial viability and reduced metabolic functions during hypothermic conditions.
Summary Insights
- Cryopreservation remains the primary cell preservation method, while hypothermic storage is gaining traction in cell therapies.
- Desiccation is a nascent alternative for long-term storage, emphasizing the need for further research.
- Understanding products' interactions with storage and transport conditions is crucial for enhancing existing processes in cell therapy.
Cell Harvesting Strategies
- Cells are detached from microcarriers through a brief period of intense agitation while in contact with a dissociation reagent.
- Concerns about cell damage during the harvesting process need to be addressed.
- SoloHill harvesting includes a 40rpm agitation with PBS wash and dissociation reagent at 37°C for 15 minutes.
- Post-dissociation, cells are retrieved using vacuum filtration.
Harvesting Efficiency
- In a 100 mL hMSC spinner flask culture after 144 hours, the actual viable cell harvest was significantly lower than expected, with efficiency below 5%.
- Comparison of harvesting methods showed a notable success rate difference: <5% efficiency for the manufacturer's method compared to >95% for the novel method.
Stress Impact on Cells
- A novel harvesting procedure utilizes higher agitation at 150rpm with a 7-minute exposure to dissociation reagent at 37°C to promote cell detachment.
- Post-harvest, cells are separated via centrifugation and vacuum filtration.
Preservation Techniques for Cell Therapy
- Different preservation techniques include desiccation, which relies on cells’ capability to withstand dehydration.
- Lyophilization (freeze-drying) removes moisture through sublimation and requires lyoprotectants, such as trehalose.
Desiccation - Lyophilization Process
- This method is critical for enabling transport at ambient temperatures, thus reducing costs.
- Freeze-drying has been implemented for various cells, including mESCs and spermatozoa, showing viable recovery of genetic material.
Research and Findings
- Studies have reported successful lyophilization of mononuclear cells from umbilical cord blood, maintaining similar levels of CD34+ HSC pre- and post-process, with 75% CFU rates.
- Viable cell recovery after lyophilization showed 90% viability, with the cells capable of proliferation and expressing vital surface markers.
Challenges in Desiccation
- Concerns exist regarding the long-term impact on cell functionality and the variance in storage conditions (temperature/humidity).
- Questions remain about the widespread application and acceptance of desiccation techniques in cell biology and regulatory environments.
Alternative Preservation Methods
- Hypothermia, or "cell pausing," involves preserving cells above 0ºC for short-term storage.
- Mild hypothermia (25-33ºC) allows cells to proliferate, which can enhance protein yield in biopharmaceutical applications.
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Description
This quiz covers essential information about the transport and handling of blood products in cell therapy. You will learn about the specific requirements for red blood cells, fresh frozen plasma, and platelets, as well as the decision factors involved in cell therapy. Understanding these aspects is crucial for effective blood product management in clinical settings.
