Human bone marrow stromal cells (hMSCs) are a promising source of multipotent progenitors for a broad range of stem cell therapies. This R03 application proposes a collaborative bioengineering research project with the objective of elucidating the mechanism of progenitor enrichment during ex vivo amplification of hMSCs at low plating density. A mechanistic understanding of the underlying population dynamics between progenitors and mature hMSCs in culture will expedite the rational design of amplification methods for the production of hMSC therapies.
The specific aim of the proposed research is to test the hypothesis that differential growth, maturation and death kinetics among progenitor and mature populations is the mechanism by which hMSC cultures at low plating density become enriched in progenitors relative to dense hMSC cultures. The hypothesis will be tested by (A) quantifying temporal changes in the accumulation of progenitor and mature hMSC populations as a function of plating density, (B) estimating kinetic parameters for hMSC population dynamics with time-lapse video microscopy, and (C) simulating population dynamics from Part A with a mathematical model using empirical data from Part B as model input. Solution of model equations will resolve the contribution of cell growth, maturation and death to progenitor enrichment. This hypothesis is based on preliminary findings by the investigators that identify progenitors in hMSC cultures and suggest an inverse relationship between plating density and progenitor content. This bioengineering project will be conducted by an interdisciplinary group of researchers with combined expertise in stem cell technology, cell population dynamics and statistical analysis, and will utilize a NIH-funded distribution center at Tulane University for adult stem cells. The integrative experimental and computational approach developed in the proposed research will serve as a template for an R01 application to improve amplification strategies for hMSCs. Results from this study can have a far-reaching impact on the progenitor content of hMSC preparations and the efficacy of hMSC therapies. (Relevance statement limited to three sentences.) Multipotent progenitors in human bone marrow stromal cells (hMSCs) exhibit a remarkable regenerative capacity that has the potential to revolutionize the treatment of damaged and diseased tissue. Collecting sufficient quantity of progenitors for clinical applications necessitates the development of ex vivo amplification methods that preserve and enrich the progenitor content of hMSCs. The proposed research will expedite the rational design of amplification methods by providing insight into the mechanism of progenitor enrichment for the case when hMSCs are cultured at a low plating density. ? ? ?
| O'Connor, Kim C; Russell, Katie C; Phinney, Donald G et al. (2011) High-capacity assay to quantify the clonal heterogeneity in potency of mesenchymal stem cells. BMC Proc 5 Suppl 8:O14 |
| Russell, Katie C; Lacey, Michelle R; Gilliam, Jennifer K et al. (2011) Clonal analysis of the proliferation potential of human bone marrow mesenchymal stem cells as a function of potency. Biotechnol Bioeng 108:2716-26 |
| Russell, Katie C; Phinney, Donald G; Lacey, Michelle R et al. (2010) In vitro high-capacity assay to quantify the clonal heterogeneity in trilineage potential of mesenchymal stem cells reveals a complex hierarchy of lineage commitment. Stem Cells 28:788-98 |
