Primary cell cultures Flashcards
What are primary cell cultures derived from?
Tissue - haemopoietic cell types or non haemopoietic.
Give examples of haemopoietic cell types?
stem cells, T and B cells, monocytes and macrophages, dendrites, neutrophils.
Give examples of non- haemopoietic cell types?
liver, muscle, skin, nerves, fibroblasts.
Outline the mechanism of dissagregaton.
cells migrate out of an explant.
Mecahnically dissociated
enzymes breakdown the cell components (trypsin, proteases and callageneases) and dissociate the cell structure.
Why do haemopoietic cells not need to be dissagegated?
They already are.
Name 3 sources of stem cells?
Red bone marrow, umbilical cord and peripheral blood.
Where is red bone marrow found in a/ adults and b/ children?
A/ - end of long bones such as the femur, humerus, skull and veterbrae.
b/ throughout the skeleton
What are the stages of maturity of a stem cell?
Progenitors, late progenitors, immature precursors (start to look different, different morphology) then distinguishable cell types.
What controls stem cell maturity growth>
Growth factors - cytokines
What does pluripotent mean in reference to early stem cells?.
Able to differentiate into any cell type.
Progenitors are committed to a particular cell lineage, how can you detect these lineages from progenitors as they are structurally the same?
Colony forming assays - allow colonies of differentiated cells to grow and see what type they are.
What are haematopoietic growth factors?
Cytokines, bind to the cell surface of transmembrane receptors and stimulate growth.
What are stromal cells?
mix of cells such as fibroblasts, macrophages, endothelial cells and adipocytes which support stem cells by giving them an environment to grow in - producing cytokines and growth factors.
What is cell processing?
Where you filter out the unwanted cells from your grown culture, to leave you with a certain cell type.
Methods of cell processing?
Antibody depletion or antibody selection.
density gradient centrifuge.
