Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture

Mirjam Fröhlich, Warren L Grayson, Darja Marolt, Jeffrey M. Gimble, Nevenka Kregar-Velikonja, Gordana Vunjak-Novakovic

Research output: Contribution to journalArticle

Abstract

We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4mm diameter×4mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400μm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-β-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.

Original languageEnglish (US)
Pages (from-to)179-189
Number of pages11
JournalTissue Engineering - Part A
Volume16
Issue number1
DOIs
StatePublished - Jan 1 2010
Externally publishedYes

Fingerprint

Bioreactors
Stem cells
Cell culture
Grafts
Bone
Stem Cells
Perfusion
Transplants
Bone and Bones
Bone Matrix
Integrin-Binding Sialoprotein
Osteopontin
Scaffolds
Tissue Engineering
Dexamethasone
Cells
Culture Media
Cell Survival
Collagen
Scaffolds (biology)

ASJC Scopus subject areas

  • Biochemistry
  • Biomedical Engineering
  • Biomaterials
  • Bioengineering
  • Medicine(all)

Cite this

Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. / Fröhlich, Mirjam; Grayson, Warren L; Marolt, Darja; Gimble, Jeffrey M.; Kregar-Velikonja, Nevenka; Vunjak-Novakovic, Gordana.

In: Tissue Engineering - Part A, Vol. 16, No. 1, 01.01.2010, p. 179-189.

Research output: Contribution to journalArticle

Fröhlich, M, Grayson, WL, Marolt, D, Gimble, JM, Kregar-Velikonja, N & Vunjak-Novakovic, G 2010, 'Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture', Tissue Engineering - Part A, vol. 16, no. 1, pp. 179-189. https://doi.org/10.1089/ten.tea.2009.0164
Fröhlich, Mirjam ; Grayson, Warren L ; Marolt, Darja ; Gimble, Jeffrey M. ; Kregar-Velikonja, Nevenka ; Vunjak-Novakovic, Gordana. / Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. In: Tissue Engineering - Part A. 2010 ; Vol. 16, No. 1. pp. 179-189.
@article{cd646d069fa54a248f89892fe144fa9b,
title = "Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture",
abstract = "We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4mm diameter×4mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400μm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-β-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.",
author = "Mirjam Fr{\"o}hlich and Grayson, {Warren L} and Darja Marolt and Gimble, {Jeffrey M.} and Nevenka Kregar-Velikonja and Gordana Vunjak-Novakovic",
year = "2010",
month = "1",
day = "1",
doi = "10.1089/ten.tea.2009.0164",
language = "English (US)",
volume = "16",
pages = "179--189",
journal = "Tissue Engineering - Part A.",
issn = "1937-3341",
publisher = "Mary Ann Liebert Inc.",
number = "1",

}

TY - JOUR

T1 - Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture

AU - Fröhlich, Mirjam

AU - Grayson, Warren L

AU - Marolt, Darja

AU - Gimble, Jeffrey M.

AU - Kregar-Velikonja, Nevenka

AU - Vunjak-Novakovic, Gordana

PY - 2010/1/1

Y1 - 2010/1/1

N2 - We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4mm diameter×4mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400μm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-β-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.

AB - We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4mm diameter×4mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400μm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-β-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.

UR - http://www.scopus.com/inward/record.url?scp=77049097956&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77049097956&partnerID=8YFLogxK

U2 - 10.1089/ten.tea.2009.0164

DO - 10.1089/ten.tea.2009.0164

M3 - Article

C2 - 19678762

AN - SCOPUS:77049097956

VL - 16

SP - 179

EP - 189

JO - Tissue Engineering - Part A.

JF - Tissue Engineering - Part A.

SN - 1937-3341

IS - 1

ER -