Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue

Szu Yu Chen, Megha Mahabole, Elan Horesh, Sara Wester, Jeffrey L. Goldberg, Scheffer C G Tseng

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

PURPOSE. Adipose-derived stem cells (ASCs) have gained importance due to their myriad potential clinical applications. We hypothesize that progenitor cells also exist besides those conventionally isolated from the stromal vascular fraction (SVF). METHOD. Central and medial orbital adipose tissues obtained from patients during eyelid surgery were digested with collagenase for 3 or 16 hours at 37°C with or without shaking. After centrifugation, the remaining cell pellet was resuspended and filtered to yield flow through in SVF and retained cells (RC) on the filter. Single cells from RC and SVF were cultured on 5% coated Matrigel in serum-free modified embryonic stem cells medium (MESCM) for 10 passages. The progenitor status was evaluated by the expression of a number of markers by qPCR and immunofluorescence staining as well as their plasticity for endothelial and tri-lineage differentiation. RESULTS. Type I collagenase digestion for 3 hours under shaking was significantly less effective in releasing progenitor cells than collagenase A digestion for 16 hours without shaking. Following filtration, cells in SVF and RC, of which the latter were tangled in collagen IVcontaining matrix, expressed different markers of progenitor cells. Cells from SVF and RC could be expanded for 10 passages on coated Matrigel in MESCM and exhibited similar or better potential to differentiate into vascular endothelial cells, chondrocytes, osteocytes, and adipocytes than SVF cells expanded on plastic in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). CONCLUSIONS. Different progenitor cells can be isolated and expanded from orbital adipose tissues. Further characterization of their mesodermal or neuroectodermal origin might enhance clinical outcome when used as a source of autologous stem cells for ocular surface regeneration.

Original languageEnglish
Pages (from-to)4842-4852
Number of pages11
JournalInvestigative Ophthalmology and Visual Science
Volume55
Issue number8
DOIs
StatePublished - Aug 6 2014

Fingerprint

Mesenchymal Stromal Cells
Blood Vessels
Adipose Tissue
Stem Cells
Stromal Cells
Matrix Metalloproteinase 13
Embryonic Stem Cells
Digestion
Osteocytes
Eagles
Collagenases
Eyelids
Chondrocytes
Serum
Centrifugation
Adipocytes
Plastics
Fluorescent Antibody Technique
Regeneration
Collagen

Keywords

  • Adipose stem cells
  • Basement membrane
  • Isolation
  • Mesenchymal stem cells
  • Orbital adipose tissue
  • Stromal vascular fraction

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue. / Chen, Szu Yu; Mahabole, Megha; Horesh, Elan; Wester, Sara; Goldberg, Jeffrey L.; Tseng, Scheffer C G.

In: Investigative Ophthalmology and Visual Science, Vol. 55, No. 8, 06.08.2014, p. 4842-4852.

Research output: Contribution to journalArticle

Chen, Szu Yu ; Mahabole, Megha ; Horesh, Elan ; Wester, Sara ; Goldberg, Jeffrey L. ; Tseng, Scheffer C G. / Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue. In: Investigative Ophthalmology and Visual Science. 2014 ; Vol. 55, No. 8. pp. 4842-4852.
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abstract = "PURPOSE. Adipose-derived stem cells (ASCs) have gained importance due to their myriad potential clinical applications. We hypothesize that progenitor cells also exist besides those conventionally isolated from the stromal vascular fraction (SVF). METHOD. Central and medial orbital adipose tissues obtained from patients during eyelid surgery were digested with collagenase for 3 or 16 hours at 37°C with or without shaking. After centrifugation, the remaining cell pellet was resuspended and filtered to yield flow through in SVF and retained cells (RC) on the filter. Single cells from RC and SVF were cultured on 5{\%} coated Matrigel in serum-free modified embryonic stem cells medium (MESCM) for 10 passages. The progenitor status was evaluated by the expression of a number of markers by qPCR and immunofluorescence staining as well as their plasticity for endothelial and tri-lineage differentiation. RESULTS. Type I collagenase digestion for 3 hours under shaking was significantly less effective in releasing progenitor cells than collagenase A digestion for 16 hours without shaking. Following filtration, cells in SVF and RC, of which the latter were tangled in collagen IVcontaining matrix, expressed different markers of progenitor cells. Cells from SVF and RC could be expanded for 10 passages on coated Matrigel in MESCM and exhibited similar or better potential to differentiate into vascular endothelial cells, chondrocytes, osteocytes, and adipocytes than SVF cells expanded on plastic in Dulbecco's modified Eagle's medium (DMEM) with 10{\%} fetal bovine serum (FBS). CONCLUSIONS. Different progenitor cells can be isolated and expanded from orbital adipose tissues. Further characterization of their mesodermal or neuroectodermal origin might enhance clinical outcome when used as a source of autologous stem cells for ocular surface regeneration.",
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T1 - Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue

AU - Chen, Szu Yu

AU - Mahabole, Megha

AU - Horesh, Elan

AU - Wester, Sara

AU - Goldberg, Jeffrey L.

AU - Tseng, Scheffer C G

PY - 2014/8/6

Y1 - 2014/8/6

N2 - PURPOSE. Adipose-derived stem cells (ASCs) have gained importance due to their myriad potential clinical applications. We hypothesize that progenitor cells also exist besides those conventionally isolated from the stromal vascular fraction (SVF). METHOD. Central and medial orbital adipose tissues obtained from patients during eyelid surgery were digested with collagenase for 3 or 16 hours at 37°C with or without shaking. After centrifugation, the remaining cell pellet was resuspended and filtered to yield flow through in SVF and retained cells (RC) on the filter. Single cells from RC and SVF were cultured on 5% coated Matrigel in serum-free modified embryonic stem cells medium (MESCM) for 10 passages. The progenitor status was evaluated by the expression of a number of markers by qPCR and immunofluorescence staining as well as their plasticity for endothelial and tri-lineage differentiation. RESULTS. Type I collagenase digestion for 3 hours under shaking was significantly less effective in releasing progenitor cells than collagenase A digestion for 16 hours without shaking. Following filtration, cells in SVF and RC, of which the latter were tangled in collagen IVcontaining matrix, expressed different markers of progenitor cells. Cells from SVF and RC could be expanded for 10 passages on coated Matrigel in MESCM and exhibited similar or better potential to differentiate into vascular endothelial cells, chondrocytes, osteocytes, and adipocytes than SVF cells expanded on plastic in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). CONCLUSIONS. Different progenitor cells can be isolated and expanded from orbital adipose tissues. Further characterization of their mesodermal or neuroectodermal origin might enhance clinical outcome when used as a source of autologous stem cells for ocular surface regeneration.

AB - PURPOSE. Adipose-derived stem cells (ASCs) have gained importance due to their myriad potential clinical applications. We hypothesize that progenitor cells also exist besides those conventionally isolated from the stromal vascular fraction (SVF). METHOD. Central and medial orbital adipose tissues obtained from patients during eyelid surgery were digested with collagenase for 3 or 16 hours at 37°C with or without shaking. After centrifugation, the remaining cell pellet was resuspended and filtered to yield flow through in SVF and retained cells (RC) on the filter. Single cells from RC and SVF were cultured on 5% coated Matrigel in serum-free modified embryonic stem cells medium (MESCM) for 10 passages. The progenitor status was evaluated by the expression of a number of markers by qPCR and immunofluorescence staining as well as their plasticity for endothelial and tri-lineage differentiation. RESULTS. Type I collagenase digestion for 3 hours under shaking was significantly less effective in releasing progenitor cells than collagenase A digestion for 16 hours without shaking. Following filtration, cells in SVF and RC, of which the latter were tangled in collagen IVcontaining matrix, expressed different markers of progenitor cells. Cells from SVF and RC could be expanded for 10 passages on coated Matrigel in MESCM and exhibited similar or better potential to differentiate into vascular endothelial cells, chondrocytes, osteocytes, and adipocytes than SVF cells expanded on plastic in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). CONCLUSIONS. Different progenitor cells can be isolated and expanded from orbital adipose tissues. Further characterization of their mesodermal or neuroectodermal origin might enhance clinical outcome when used as a source of autologous stem cells for ocular surface regeneration.

KW - Adipose stem cells

KW - Basement membrane

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KW - Orbital adipose tissue

KW - Stromal vascular fraction

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