Multifactorial Experimental Design to Optimize the Anti-Inflammatory and Proangiogenic Potential of Mesenchymal Stem Cell Spheroids

Kaitlin C. Murphy, Jacklyn Whitehead, Patrick C. Falahee, Dejie Zhou, Scott I. Simon, Jonathan K Leach

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Mesenchymal stem cell therapies promote wound healing by manipulating the local environment to enhance the function of host cells. Aggregation of mesenchymal stem cells (MSCs) into three-dimensional spheroids increases cell survival and augments their anti-inflammatory and proangiogenic potential, yet there is no consensus on the preferred conditions for maximizing spheroid function in this application. The objective of this study was to optimize conditions for forming MSC spheroids that simultaneously enhance their anti-inflammatory and proangiogenic nature. We applied a design of experiments (DOE) approach to determine the interaction between three input variables (number of cells per spheroid, oxygen tension, and inflammatory stimulus) on MSC spheroids by quantifying secretion of prostaglandin E2 (PGE2) and vascular endothelial growth factor (VEGF), two potent molecules in the MSC secretome. DOE results revealed that MSC spheroids formed with 40,000 cells per spheroid in 1% oxygen with an inflammatory stimulus (Spheroid 1) would exhibit enhanced PGE2 and VEGF production versus those formed with 10,000 cells per spheroid in 21% oxygen with no inflammatory stimulus (Spheroid 2). Compared to Spheroid 2, Spheroid 1 produced fivefold more PGE2 and fourfold more VEGF, providing the opportunity to simultaneously upregulate the secretion of these factors from the same spheroid. The spheroids induced macrophage polarization, sprout formation with endothelial cells, and keratinocyte migration in a human skin equivalent model-demonstrating efficacy on three key cell types that are dysfunctional in chronic non-healing wounds. We conclude that DOE-based analysis effectively identifies optimal culture conditions to enhance the anti-inflammatory and proangiogenic potential of MSC spheroids. Stem Cells 2017

Original languageEnglish (US)
JournalStem Cells
DOIs
StateAccepted/In press - 2017

Fingerprint

Mesenchymal Stromal Cells
Anti-Inflammatory Agents
Research Design
Dinoprostone
Vascular Endothelial Growth Factor A
Oxygen
Cell- and Tissue-Based Therapy
Keratinocytes
Wound Healing
Cell Movement
Cell Survival
Up-Regulation
Stem Cells
Endothelial Cells
Cell Count
Macrophages
Skin
Wounds and Injuries

Keywords

  • Design of experiments
  • Inflammation
  • Mesenchymal stem cell
  • Proangiogenic
  • Prostaglandin E2

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Cite this

Multifactorial Experimental Design to Optimize the Anti-Inflammatory and Proangiogenic Potential of Mesenchymal Stem Cell Spheroids. / Murphy, Kaitlin C.; Whitehead, Jacklyn; Falahee, Patrick C.; Zhou, Dejie; Simon, Scott I.; Leach, Jonathan K.

In: Stem Cells, 2017.

Research output: Contribution to journalArticle

Murphy, Kaitlin C. ; Whitehead, Jacklyn ; Falahee, Patrick C. ; Zhou, Dejie ; Simon, Scott I. ; Leach, Jonathan K. / Multifactorial Experimental Design to Optimize the Anti-Inflammatory and Proangiogenic Potential of Mesenchymal Stem Cell Spheroids. In: Stem Cells. 2017.
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