Abstract
Stem cells from the eye hold a great potential for vision restoration and can also be used for regeneration in other tissues. In this study, we characterized the stem cell properties of Trabecular meshwork stem cells (TMSCs) after long-term cryopreservation (∼8 years). TMSCs derived from four donors were examined for their viability and proliferation, as well as stem cell marker expression. Spheroid formation, colony formation, and multipotency were investigated. We observed that TMSCs were fully viable with variable proliferation ability. They expressed the stem cell markers CD90, CD166, CD105, CD73, OCT4, SSEA4, Notch1, KLF4, ABCG2, Nestin, and HNK1 detected by flow cytometry, quantitative polymerase chain reaction, or immunofluorescent staining. They could form spheroids and colonies after thawing. All TMSCs were able to differentiate into osteocytes, neural cells, and trabecular meshwork (TM) cells, but not adipocytes. Differentiated TM cells responded to dexamethasone treatment with increased expression of myocilin and angiopoietin-like 7 (ANGPTL7). In a nutshell, our study demonstrated that TMSCs retain their stem cell properties after long-term cryopreservation and hence can be an effective cell therapy source for various clinical applications.
Keywords:
cryopreservation; multipotency; regeneration; stem cells; trabecular meshwork.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Angiopoietin-Like Protein 7
-
Angiopoietin-like Proteins / genetics
-
Angiopoietin-like Proteins / metabolism
-
Anti-Inflammatory Agents / pharmacology
-
Cell Differentiation / genetics*
-
Cell Lineage / genetics
-
Cells, Cultured
-
Cryopreservation / methods*
-
Cytoskeletal Proteins / genetics
-
Cytoskeletal Proteins / metabolism
-
Dexamethasone / pharmacology
-
Eye Proteins / genetics
-
Eye Proteins / metabolism
-
Flow Cytometry
-
Glycoproteins / genetics
-
Glycoproteins / metabolism
-
Humans
-
Kruppel-Like Factor 4
-
Kruppel-Like Transcription Factors / genetics
-
Kruppel-Like Transcription Factors / metabolism
-
Neurons / cytology
-
Neurons / metabolism
-
Octamer Transcription Factor-3 / genetics
-
Octamer Transcription Factor-3 / metabolism
-
Osteocytes / cytology
-
Osteocytes / metabolism
-
Reverse Transcriptase Polymerase Chain Reaction
-
Stem Cells / cytology
-
Stem Cells / drug effects
-
Stem Cells / metabolism*
-
Time Factors
-
Trabecular Meshwork / cytology
-
Trabecular Meshwork / metabolism*
Substances
-
ANGPTL7 protein, human
-
Angiopoietin-Like Protein 7
-
Angiopoietin-like Proteins
-
Anti-Inflammatory Agents
-
Cytoskeletal Proteins
-
Eye Proteins
-
Glycoproteins
-
KLF4 protein, human
-
Kruppel-Like Factor 4
-
Kruppel-Like Transcription Factors
-
Octamer Transcription Factor-3
-
POU5F1 protein, human
-
trabecular meshwork-induced glucocorticoid response protein
-
Dexamethasone