text-only page produced automatically by Usablenet Assistive Skip all navigation and go to page content Skip top navigation and go to directorate navigation Skip top navigation and go to page navigation
National Science Foundation
Awards
design element
Search Awards
Recent Awards
Presidential and Honorary Awards
About Awards
Grant Policy Manual
Grant General Conditions
Cooperative Agreement Conditions
Special Conditions
Federal Demonstration Partnership
Policy Office Website



Award Abstract #1253189

CAREER: Mechanical Biomarkers and Mesenchymal Stem Cell Differentiation

NSF Org: CBET
Div Of Chem, Bioeng, Env, & Transp Sys
divider line
Initial Amendment Date: February 1, 2013
divider line
Latest Amendment Date: March 4, 2016
divider line
Award Number: 1253189
divider line
Award Instrument: Standard Grant
divider line
Program Manager: Carol Lucas
CBET Div Of Chem, Bioeng, Env, & Transp Sys
ENG Directorate For Engineering
divider line
Start Date: February 1, 2013
divider line
End Date: January 31, 2018 (Estimated)
divider line
Awarded Amount to Date: $437,792.00
divider line
Investigator(s): Eric Darling Eric_Darling@brown.edu (Principal Investigator)
divider line
Sponsor: Brown University
BOX 1929
Providence, RI 02912-9002 (401)863-2777
divider line
NSF Program(s): BIOMEDICAL ENGINEERING,
EXP PROG TO STIM COMP RES
divider line
Program Reference Code(s): 004E, 017E, 1045, 138E, 7218, 9150
divider line
Program Element Code(s): 5345, 9150

ABSTRACT

1253189

Darling

Mesenchymal stem cells (MSCs) pose exciting possibilities for repairing tissues damaged by injury or disease. Their relative abundance and capability to become many different cell types make them attractive as an alternative autologous cell source. However, the cellular heterogeneity present in MSC harvests complicates their therapeutic application since not all cells will behave in the same manner. Individual stem cells respond to biochemical and mechanical stimuli in their local microenvironment, which can direct differentiation along specific pathways/lineages. While substrate compliance has been extensively investigated in recent years as it relates to stem cell differentiation, little attention has been given to how the mechanical properties of individual cells can influence this process for themselves and their neighbors. Findings from the PI?s laboratory indicate cellular mechanical properties correlate with biological characteristics, such as gene expressions, and play an important role in determining the differentiation fate of stem cells. Not all MSCs behave uniformly, and this heterogeneity can dramatically impact the overall response of a sample. Understanding how the biological and mechanical characteristics of local cell populations relate to the larger group can provide clues to optimizing future regenerative therapies.

The proposed project will investigate the heterogeneity of MSC samples at the single-cell, neighboring (~101 cells), and population (~103+ cells) levels using two complimentary strategies. First, live-cell, gene expression-based markers will be used to identify differentiating and non-differentiating MSCs that have been chemically induced for osteogenesis and adipogenesis. Second, single-cell, elastic and viscoelastic testing will assess the mechanical heterogeneity in undifferentiated, ?partially? differentiated, and fully differentiated stem cell cultures. In addition to expanding knowledge of stem cell mechanics, this research program has the potential to vastly improve experimental approaches that strive to direct stem cell differentiation and improve regenerative responses following implantation.

Intellectual Merit: The proposed project seeks to investigate the inherent heterogeneity present in mesenchymal stem cell populations. MSCs are typically investigated at the population level, which can obscure variations that exist among individual cells. Little is known about how single-cell, mechanical properties change during differentiation, but this knowledge is critical for fully understanding cell-substrate and cell-cell behavior. Likewise, elucidating the spatiotemporal patterns of gene expression in MSC samples can help determine optimal times and locations for biochemical stimulation. To pursue these goals, this project will use custom-designed molecular beacons to visualize mRNA molecules in live, differentiating MSCs. Atomic force microscopy will be used to mechanically characterize the elastic and viscoelastic properties of individual cells. Focus will be given to how local cell populations behave, biochemically and mechanically, in relation to the overall sample. Findings will be applicable to many fields, including stem cell biology, cell mechanics, and tissue engineering. The PI is a pioneer in the area of single-cell, mechanical biomarkers, an exciting new field that provides phenotypic characterization akin to gene and protein expression. The proposed research furthers the maturation of these transformative approaches by investigating the role of mechanical biomarkers in stem cell differentiation.

Broader Impact: As a biomedical engineer conducting research at the cross-section of math, biology, chemistry, and engineering, the PI is well-suited to provide an environment that highlights the need for interdisciplinary understanding. The outreach effort proposed here will provide primary and secondary school educators at the Providence Public School District the opportunity to conduct team-based research during the summer. Teachers specializing in different subjects will work together on projects that incorporate multiple areas of expertise. The goal of this project is to facilitate interaction among teachers so that they can develop interdisciplinary examples and experiments for their own classes. Students take many math and science courses throughout their education, encompassing algebra, geometry, biology, chemistry, and physics. However, these courses are almost always taught as isolated subjects, whereas real-world applications are increasingly interdisciplinary. The proposed project will be conducted as an extension of Brown University?s established GK-12 program. In addition to this outreach effort, the PI will refine his recently designed course on stem cell engineering for graduate and undergraduate students that covers areas of stem cell biology, regenerative medicine, and hands-on laboratory exercises. Experimental findings from the research portion of this proposal will be directly applicable to this course, which is the first of its kind at Brown University.


PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

Note:  When clicking on a Digital Object Identifier (DOI) number, you will be taken to an external site maintained by the publisher. Some full text articles may not yet be available without a charge during the embargo (administrative interval).

Some links on this page may take you to non-federal websites. Their policies may differ from this site.


Kanthilal M, Darling EM. "Characterization of mechanical and regenerative properties of human, adipose stromal cells," Cell Mol Bioeng, v.7, 2014, p. 585.

Beane OS, Fonseca VC, Darling EM.. "Adipose-derived stem cells retain their regenerative potential after methotrexate treatment," Exp Cell Res, v.327, 2014, p. 222.

Marble HD, Sutermaster BA, Kanthilal M, Fonseca VC, Darling EM. "Gene expression-based enrichment of live cells from adipose tissue produces subpopulations with improved osteogenic potential," Stem Cell Res Ther, v.5, 2014, p. 12.

Labriola NR, Darling EM. "Temporal heterogeneity in single-cell gene expression and mechanical properties during adipogenic differentiation," J Biomech, v.48, 2015.

Dingle YL, Chirila AM, Boutin ME, Livi LL, Labriola NR, Jakubek LM, Morgan JR, Darling EM, Kauer JA, Hoffman-Kim D. "3D neural spheroid culture: An in vitro model for the central nervous system," Tissue Eng A, 2015.

 

Please report errors in award information by writing to: awardsearch@nsf.gov.

 

 

Print this page
Back to Top of page
  FUNDING   AWARDS   DISCOVERIES   NEWS   PUBLICATIONS   STATISTICS   ABOUT NSF   FASTLANE  
Research.gov  |  USA.gov  |  National Science Board  |  Recovery Act  |  Budget and Performance  |  Annual Financial Report
Web Policies and Important Links  |  Privacy  |  FOIA  |  NO FEAR Act  |  Inspector General  |  Webmaster Contact  |  Site Map
National Science Foundation Logo
The National Science Foundation, 4201 Wilson Boulevard, Arlington, Virginia 22230, USA
Tel: (703) 292-5111, FIRS: (800) 877-8339 | TDD: (800) 281-8749
  Text Only Version