Journal article
Effects of plating density and culture time on bone marrow stromal cell characteristics
Experimental hematology, v 36(9), pp 1176-1185
01 Sep 2008
PMID: 18495329
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Objective. Bone marrow stromal cells (MSC) are multipotent adult stem cells that have emerged as promising candidates for cell therapy in disorders including cardiac infarction, stroke, and spinal cord injury. While harvesting methods used by different laboratories are relatively standard, MSC culturing protocols vary widely. This study is aimed at evaluating the effects of initial plating density and total time in culture on proliferation, cell morphology, and differentiation potential of heterogeneous MSC cultures and more homogeneous cloned subpopulations.
Materials and Methods. Rat MSC were plated at 20, 200, and 2000 cells/cm(2) and grown to 50% confluency. The numbers of population doublings and doubling times were determined within and across multiple passages. Changes in cell morphology and differentiation potential to adipogenic, chondrogenic, and osteogenic lineages were evaluated and compared among early, intermediate, and late passages, as well as between heterogeneous and cloned MSC Populations.
Results. We found optimal cell growth at a plating density of 200 cells/cm(2). Cultures derived from all plating densities developed increased proportions of flat cells over time. Assays for chondrogenesis, osteogenesis, and adipogenesis showed that heterogeneous MSC plated at all densities sustained the potential for all three mesenchymal phenotypes through at least passage 5; the flat subpopulation lost adipogenic and chondrogenic potential.
Conclusion. Our findings suggest that the initial plating density is not critical for maintaining a well-defined, multipotent MSC population. Time in culture, however, affects cell characteristics, suggesting that cell expansion should be limited, especially until the specific characteristics of different MSC subpopulations are better understood. (C) 2008 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.
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Details
- Title
- Effects of plating density and culture time on bone marrow stromal cell characteristics
- Creators
- Birgit Neuhuber - Drexel UniversitySharon A. Swanger - Drexel UniversityLinda Howard - Osiris TherapeuticsAlastair Mackay - Osiris TherapeuticsItzhak Fischer - Drexel University
- Publication Details
- Experimental hematology, v 36(9), pp 1176-1185
- Publisher
- Elsevier
- Number of pages
- 10
- Grant note
- 70NAN2H3017 / National Institute of Standards and Technology; National Institute of Standards & Technology (NIST) - USA 8570 / Shriners Hospital NS 43882; NS 049429 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA R21NS049429 / NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Neurological Disorders & Stroke (NINDS)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Neurobiology and Anatomy
- Web of Science ID
- WOS:000258943100013
- Scopus ID
- 2-s2.0-49349108165
- Other Identifier
- 991019168325104721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Hematology
- Medicine, Research & Experimental