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Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue.
Invest Ophthalmol Vis Sci 2014; 55(8):4842-52IO

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 IV-containing 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.

Authors+Show Affiliations

TissueTech, Inc., Miami, Florida, United States.TissueTech, Inc., Miami, Florida, United States.University of Miami Miller School of Medicine, Miami, Florida, United States.University of Miami Miller School of Medicine, Miami, Florida, United States Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States.University of Miami Miller School of Medicine, Miami, Florida, United States.TissueTech, Inc., Miami, Florida, United States Ocular Surface Center, and Ocular Surface Research Education Foundation, Miami, Florida, United States.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24994870

Citation

Chen, Szu-Yu, et al. "Isolation and Characterization of Mesenchymal Progenitor Cells From Human Orbital Adipose Tissue." Investigative Ophthalmology & Visual Science, vol. 55, no. 8, 2014, pp. 4842-52.
Chen SY, Mahabole M, Horesh E, et al. Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue. Invest Ophthalmol Vis Sci. 2014;55(8):4842-52.
Chen, S. Y., Mahabole, M., Horesh, E., Wester, S., Goldberg, J. L., & Tseng, S. C. (2014). Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue. Investigative Ophthalmology & Visual Science, 55(8), pp. 4842-52. doi:10.1167/iovs.14-14441.
Chen SY, et al. Isolation and Characterization of Mesenchymal Progenitor Cells From Human Orbital Adipose Tissue. Invest Ophthalmol Vis Sci. 2014 Jul 3;55(8):4842-52. PubMed PMID: 24994870.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR 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, Y1 - 2014/07/03/ PY - 2014/7/5/entrez PY - 2014/7/6/pubmed PY - 2014/10/15/medline KW - adipose stem cells KW - basement membrane KW - isolation KW - mesenchymal stem cells KW - orbital adipose tissue KW - stromal vascular fraction SP - 4842 EP - 52 JF - Investigative ophthalmology & visual science JO - Invest. Ophthalmol. Vis. Sci. VL - 55 IS - 8 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 IV-containing 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. SN - 1552-5783 UR - https://www.unboundmedicine.com/medline/citation/24994870/Isolation_and_characterization_of_mesenchymal_progenitor_cells_from_human_orbital_adipose_tissue_ L2 - http://iovs.arvojournals.org/article.aspx?doi=10.1167/iovs.14-14441 DB - PRIME DP - Unbound Medicine ER -