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  • Human Schwann Cells in vitro III. Analytical Methods and a Practical Approach for Quality Control.

Human Schwann Cells in vitro III. Analytical Methods and a Practical Approach for Quality Control.

Bio-protocol (2023-11-30)
Paula V Monje
ABSTRACT

This paper introduces simple analytical methods and bioassays to promptly assess the identity and function of in vitro cultured human Schwann cells (hSCs). A systematic approach is proposed to unequivocally discriminate hSCs from other glial cells, non-glial cells, and non-human SCs (authentication), identify hSCs at different stages of differentiation, and determine whether individual hSCs are proliferative or senescent. Examples of how to use distinct cell-based approaches for quality control and routine troubleshooting are provided to confirm the constitution (identity, purity, and heterogeneity) and potency (bioactivity) of hSC cultures from multiple sources. The bioassays are valuable for rapidly gauging the responses of hSCs to mitogenic and differentiating factors and ascertaining the cells' basic properties before performing co-culture or cell grafting studies. The assays are image based and use adherent hSCs established in monoculture to simplify the experimental setup and interpretation of results. Finally, all sections contain thorough background information, notes, and references to facilitate decision making, data interpretation, and ad hoc method development for diverse applications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Po Antibody, Chemicon®, from chicken
Sigma-Aldrich
Anti-Myelin Basic Protein Antibody, a.a. 82-87, culture supernatant, clone 12, Chemicon®
Sigma-Aldrich
Adenosine 3′,5′-cyclic Monophosphate, 8-(4-Chlorophenylthio)-, Sodium Salt, Cell-permeable cAMP analog that activates both protein kinase A and protein kinase G.
Sigma-Aldrich
Anti-Nuclei Antibody, clone 235-1, clone 235-1, Chemicon®, from mouse