TY  - JOUR
TI  - Human Schwann Cells in vitro II. Passaging, Purification, Banking, and Labeling of Established Cultures
AU  - Monje, Paula V
VL  - 13
IS  - 22
PY  - 2023
DA  - 2023/11/20
SP  - e4882
C1  - Bio-protocol 2023;13:e4882
DO  - 10.21769/BioProtoc.4882
UR  - https://doi.org/10.21769/BioProtoc.4882
AB  - This manuscript describes step-by-step procedures to establish and manage fresh and cryopreserved cultures of nerve-derived human Schwann cells (hSCs) at the desired scale. Adaptable protocols are provided to propagate hSC cultures through serial passaging and perform routine manipulations such as enzymatic dissociation, purification, cryogenic preservation, live-cell labeling, and gene delivery. Expanded hSCs cultures are metabolically active, proliferative, and phenotypically stable for at least three consecutive passages. Cell yields are expected to be variable as determined by the rate of growth of individual batches and the rounds of subculture. The purity, however, can be maintained high at >95% hSC regardless of passage. The cells obtained in this manner are suitable for various applications, including small drug screens, in vitro modeling of neurodevelopmental processes, and cell transplantation. One caveat of this protocol is that continued expansion of same-batch hSC populations is eventually restricted due to senescence-linked growth arrest.
KW  - Ensheathing glia
KW  - Peripheral nerve
KW  - Cell culture
KW  - Mitogenic factors
KW  - Laminin
KW  - Serial passaging
KW  - Fibroblast contamination
KW  - Immunopanning
KW  - Magnetic-activated cell sorting
KW  - Cryopreservation
KW  - Scalability
KW  - Proliferation
KW  - Senescence
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False