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Propagation and senescence of human marrow stromal cells in culture: a simple colony‐forming assay identifies samples with the greatest potential to propagate and differentiate
Author(s) -
DiGirolamo Carla M.,
Stokes David,
Colter David,
Phinney Donald G.,
Class Reiner,
Prockop Darwin J.
Publication year - 1999
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1999.01715.x
Subject(s) - stromal cell , bone marrow , biology , mesenchymal stem cell , cell culture , serial passage , colony forming unit , iliac crest , stem cell , tissue culture , cell , microbiology and biotechnology , senescence , immunology , andrology , in vitro , cancer research , genetics , medicine , anatomy , bacteria
Marrow stromal cells (MSCs) were isolated from bone marrow obtained by aspirates of the iliac crest of normal volunteers. The cells were isolated by their adherence to plastic and then passed in culture. Some of the samples expanded through over 15 cell doublings from the time frozen stocks were prepared. Others ceased replicating after about four cell doublings. The replicative potential of the cells in culture was best predicted by a simple colony‐forming assay in which samples from early passages were plated at low densities of about 10 cells per cm 2 . Samples with high colony‐forming efficiency exhibited the greatest replicative potential. The colonies obtained by plating early passage cells at low density varied in size and morphology. The large colonies readily differentiated into osteoblasts and adipocytes when incubated in the appropriate medium. As samples were expanded in culture and approached senescence, they retained their ability to differentiate into osteoblasts. However, the cells failed to differentiate into adipocytes. The loss of multipotentiality following serial passage in culture may have important implications for the use of expanded MSCs for cell and gene therapy.