Human mesenchymal stem cell spheroids in fibrin hydrogels exhibit improved cell survival and potential for bone healing

Kaitlin C. Murphy, Sophia Y Fang, Jonathan K Leach

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Mesenchymal stem cells (MSCs) have great therapeutic potential for the repair of nonhealing bone defects, because of their proliferative capacity, multilineage potential, trophic factor secretion and lack of immunogenicity. However, a major challenge to the translation of cell-based therapies into clinical practice is ensuring their survival and function upon implantation into the defect site. We hypothesize that forming MSCs into more physiologic three-dimensional spheroids, rather than employing dissociated cells from two-dimensional monolayer culture, will enhance their survival when exposed to a harsh microenvironment but maintain their osteogenic potential. MSC spheroids were formed by using the hanging drop method with increasing cell numbers. Compared with larger spheroids, the smallest spheroids, which contained 15,000 cells, exhibited increased metabolic activity, reduced apoptosis and the most uniform distribution of proliferating cells. Spheroids were then entrapped in fibrin gels and cultured in serum-free medium and 1 % oxygen. Compared with identical numbers of dissociated MSCs in fibrin gels, spheroids exhibited significantly reduced apoptosis and secreted up to 100-fold more vascular endothelial growth factor. Moreover, fibrin gels containing spheroids and those containing an equivalent number of dissociated cells exhibited similar expression levels of early and late markers of osteogenic differentiation. Thus, MSC spheroids exhibit greater resistance to apoptosis and enhanced proangiogenic potential while maintaining similar osteogenic potential to dissociated MSCs entrapped in a clinically relevant biomaterial, supporting the use of MSC spheroids in cell-based approaches to bone repair.

Original languageEnglish (US)
Pages (from-to)91-99
Number of pages9
JournalCell and Tissue Research
Volume357
Issue number1
DOIs
StatePublished - 2014

Fingerprint

Hydrogels
Fibrin
Mesenchymal Stromal Cells
Cell Survival
Bone and Bones
Gels
Apoptosis
Cell Count
Survival
Differentiation Antigens
Serum-Free Culture Media
Biocompatible Materials
Cell- and Tissue-Based Therapy
Vascular Endothelial Growth Factor A
Oxygen

Keywords

  • Cell survival
  • Fibrin
  • Mesenchymal stem cell
  • Osteogenic potential
  • Spheroid

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Histology

Cite this

Human mesenchymal stem cell spheroids in fibrin hydrogels exhibit improved cell survival and potential for bone healing. / Murphy, Kaitlin C.; Fang, Sophia Y; Leach, Jonathan K.

In: Cell and Tissue Research, Vol. 357, No. 1, 2014, p. 91-99.

Research output: Contribution to journalArticle

Murphy, Kaitlin C. ; Fang, Sophia Y ; Leach, Jonathan K. / Human mesenchymal stem cell spheroids in fibrin hydrogels exhibit improved cell survival and potential for bone healing. In: Cell and Tissue Research. 2014 ; Vol. 357, No. 1. pp. 91-99.
@article{9246c4b234be4815b43d35eaa022ee1f,
title = "Human mesenchymal stem cell spheroids in fibrin hydrogels exhibit improved cell survival and potential for bone healing",
abstract = "Mesenchymal stem cells (MSCs) have great therapeutic potential for the repair of nonhealing bone defects, because of their proliferative capacity, multilineage potential, trophic factor secretion and lack of immunogenicity. However, a major challenge to the translation of cell-based therapies into clinical practice is ensuring their survival and function upon implantation into the defect site. We hypothesize that forming MSCs into more physiologic three-dimensional spheroids, rather than employing dissociated cells from two-dimensional monolayer culture, will enhance their survival when exposed to a harsh microenvironment but maintain their osteogenic potential. MSC spheroids were formed by using the hanging drop method with increasing cell numbers. Compared with larger spheroids, the smallest spheroids, which contained 15,000 cells, exhibited increased metabolic activity, reduced apoptosis and the most uniform distribution of proliferating cells. Spheroids were then entrapped in fibrin gels and cultured in serum-free medium and 1 {\%} oxygen. Compared with identical numbers of dissociated MSCs in fibrin gels, spheroids exhibited significantly reduced apoptosis and secreted up to 100-fold more vascular endothelial growth factor. Moreover, fibrin gels containing spheroids and those containing an equivalent number of dissociated cells exhibited similar expression levels of early and late markers of osteogenic differentiation. Thus, MSC spheroids exhibit greater resistance to apoptosis and enhanced proangiogenic potential while maintaining similar osteogenic potential to dissociated MSCs entrapped in a clinically relevant biomaterial, supporting the use of MSC spheroids in cell-based approaches to bone repair.",
keywords = "Cell survival, Fibrin, Mesenchymal stem cell, Osteogenic potential, Spheroid",
author = "Murphy, {Kaitlin C.} and Fang, {Sophia Y} and Leach, {Jonathan K}",
year = "2014",
doi = "10.1007/s00441-014-1830-z",
language = "English (US)",
volume = "357",
pages = "91--99",
journal = "Cell and Tissue Research",
issn = "0302-766X",
publisher = "Springer Verlag",
number = "1",

}

TY - JOUR

T1 - Human mesenchymal stem cell spheroids in fibrin hydrogels exhibit improved cell survival and potential for bone healing

AU - Murphy, Kaitlin C.

AU - Fang, Sophia Y

AU - Leach, Jonathan K

PY - 2014

Y1 - 2014

N2 - Mesenchymal stem cells (MSCs) have great therapeutic potential for the repair of nonhealing bone defects, because of their proliferative capacity, multilineage potential, trophic factor secretion and lack of immunogenicity. However, a major challenge to the translation of cell-based therapies into clinical practice is ensuring their survival and function upon implantation into the defect site. We hypothesize that forming MSCs into more physiologic three-dimensional spheroids, rather than employing dissociated cells from two-dimensional monolayer culture, will enhance their survival when exposed to a harsh microenvironment but maintain their osteogenic potential. MSC spheroids were formed by using the hanging drop method with increasing cell numbers. Compared with larger spheroids, the smallest spheroids, which contained 15,000 cells, exhibited increased metabolic activity, reduced apoptosis and the most uniform distribution of proliferating cells. Spheroids were then entrapped in fibrin gels and cultured in serum-free medium and 1 % oxygen. Compared with identical numbers of dissociated MSCs in fibrin gels, spheroids exhibited significantly reduced apoptosis and secreted up to 100-fold more vascular endothelial growth factor. Moreover, fibrin gels containing spheroids and those containing an equivalent number of dissociated cells exhibited similar expression levels of early and late markers of osteogenic differentiation. Thus, MSC spheroids exhibit greater resistance to apoptosis and enhanced proangiogenic potential while maintaining similar osteogenic potential to dissociated MSCs entrapped in a clinically relevant biomaterial, supporting the use of MSC spheroids in cell-based approaches to bone repair.

AB - Mesenchymal stem cells (MSCs) have great therapeutic potential for the repair of nonhealing bone defects, because of their proliferative capacity, multilineage potential, trophic factor secretion and lack of immunogenicity. However, a major challenge to the translation of cell-based therapies into clinical practice is ensuring their survival and function upon implantation into the defect site. We hypothesize that forming MSCs into more physiologic three-dimensional spheroids, rather than employing dissociated cells from two-dimensional monolayer culture, will enhance their survival when exposed to a harsh microenvironment but maintain their osteogenic potential. MSC spheroids were formed by using the hanging drop method with increasing cell numbers. Compared with larger spheroids, the smallest spheroids, which contained 15,000 cells, exhibited increased metabolic activity, reduced apoptosis and the most uniform distribution of proliferating cells. Spheroids were then entrapped in fibrin gels and cultured in serum-free medium and 1 % oxygen. Compared with identical numbers of dissociated MSCs in fibrin gels, spheroids exhibited significantly reduced apoptosis and secreted up to 100-fold more vascular endothelial growth factor. Moreover, fibrin gels containing spheroids and those containing an equivalent number of dissociated cells exhibited similar expression levels of early and late markers of osteogenic differentiation. Thus, MSC spheroids exhibit greater resistance to apoptosis and enhanced proangiogenic potential while maintaining similar osteogenic potential to dissociated MSCs entrapped in a clinically relevant biomaterial, supporting the use of MSC spheroids in cell-based approaches to bone repair.

KW - Cell survival

KW - Fibrin

KW - Mesenchymal stem cell

KW - Osteogenic potential

KW - Spheroid

UR - http://www.scopus.com/inward/record.url?scp=84904676373&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84904676373&partnerID=8YFLogxK

U2 - 10.1007/s00441-014-1830-z

DO - 10.1007/s00441-014-1830-z

M3 - Article

C2 - 24781147

AN - SCOPUS:84904676373

VL - 357

SP - 91

EP - 99

JO - Cell and Tissue Research

JF - Cell and Tissue Research

SN - 0302-766X

IS - 1

ER -