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Cell Banking of hiPSCs: A Practical Guide to Cryopreservation and Quality Control in Basic Research
Aya Shibamiya1,2, Elisabeth Schulze1,2, Dana Krauß1,3
1Institute of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Insights
Establishing quality-controlled human induced pluripotent stem cell (hiPSC) banks ensures research reproducibility. This study details a cost-effective method for generating hiPSC master and working cell banks with essential quality assurance protocols.
Area of Science:
- Stem Cell Biology
- Cell Banking
- Research Reproducibility
Background:
- Stem cell research reproducibility depends on consistent access to high-quality cells.
- Human induced pluripotent stem cells (hiPSCs) can degrade over passages, impacting experimental outcomes.
- Cell banking is crucial for maintaining hiPSC quality and minimizing batch-to-batch variability.
Purpose of the Study:
- To present an economical strategy for creating master and working cell banks of hiPSCs for basic research.
- To outline essential quality assurance (QA) protocols for banked hiPSCs.
- To ensure reliable and consistent results in stem cell research.
Main Methods:
- Development of cost-efficient protocols for hiPSC expansion and cell banking.
- Implementation of minimal QA tests including sterility, viability, pluripotency, and genetic integrity assessments.
- Detailed protocols for cell banking (Basic Protocol 2) and quality control.
Main Results:
- A defined, cost-effective method for generating hiPSC master and working cell banks was established.
- Minimal QA protocols were described, covering critical aspects of cell quality.
- The protocols support consistent cell availability for reproducible stem cell research.
Conclusions:
- Implementing standardized cell banking and QA procedures is vital for reproducible stem cell research.
- This work provides a practical and economical approach for researchers to establish reliable hiPSC stocks.
- Consistent quality of hiPSCs through banking ensures the validity and comparability of research findings.
Abstract:
The reproducibility of stem cell research relies on the constant availability of quality-controlled cells. As the quality of human induced pluripotent stem cells (hiPSCs) can deteriorate in the course of a few passages, cell banking is key to achieve consistent results and low batch-to-batch variation. Here, we provide a cost-efficient route to generate master and working cell banks for basic research projects. In addition, we describe minimal protocols for quality assurance including tests for sterility, viability, pluripotency, and genetic integrity. © 2020 The Authors. Basic Protocol 1: Expansion of hiPSCs Basic Protocol 2: Cell banking of hiPSCs Support Protocol 1: Pluripotency assessment by flow cytometry Support Protocol 2: Thawing control: Viability and sterility Support Protocol 3: Potency, viral clearance, and pluripotency: Spontaneous differentiation and qRT-PCR Support Protocol 4: Identity: Short tandem repeat analysis.
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