Premium
Development of quantitative PCR methods to analyse neural progenitor cell culture state
Author(s) -
Abranches Elsa,
O'Neill Analeah,
Robertson Matthew J.,
Schaffer David V.,
Cabral Joaquim M. S.
Publication year - 2006
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20050218
Subject(s) - progenitor cell , biology , stem cell , neural stem cell , microbiology and biotechnology , cellular differentiation , cell culture , regeneration (biology) , progenitor , immunology , computational biology , genetics , gene
Stem cells have significant potential for tissue engineering and regeneration, and neural stem and progenitor cells have proven promising for neuroregeneration in numerous animal disease and injury models. However, improved approaches must be developed to culture, expand and control the cells. Therefore the development of enhanced methods to quantify cell differentiation would significantly aid both in the basic investigation of cell‐fate control mechanisms and in the optimization and validation of cell culture and expansion conditions. Quantitative reverse transcription–PCR methods were developed to quantify cell differentiation state by monitoring the expression of several cell‐lineage‐specific markers. These methods provide more rapid and readily quantitative results when compared with immunostaining. These methods were also applied in a preliminary investigation of cell‐culture conditions, and it was found that regular feeding of cells with fresh medium is necessary to maintain them in an undifferentiated and highly proliferative state. The present study may aid both basic efforts to study the control of neural stem and progenitor differentiation as well as endeavours to optimize cell culture and expansion conditions for biomedical applications.