Characterizing differences between MSCs and TM cells: Toward autologous stem cell therapies for the glaucomatous trabecular meshwork.
Eric J SniderR Taylor VannattaLisa SchildmeyerW Daniel StamerC Ross EthierPublished in: Journal of tissue engineering and regenerative medicine (2017)
Glaucoma, a leading cause of blindness, is characterized by an increase in intraocular pressure, which is largely determined by resistance to aqueous humour outflow through the trabecular meshwork (TM). In glaucoma, the cellularity of the TM is decreased, and, as a result, stem cell therapies for the TM represent a potential therapeutic option for restoring TM function and treating glaucoma patients. We here focus on adipose derived mesenchymal stem cells (MSCs) as a potential autologous cell source for TM regenerative medicine applications and describe characterization techniques at the messenger (reverse transcription-quantitative polymerase chain reaction), protein (western blotting, flow cytometry), and functional (contractility, phagocytosis) levels to distinguish MSCs from TM cells. We present a panel of 12 transcripts to allow: (a) suitable normalization of reverse transcription-quantitative polymerase chain reaction results across cell types and after exposure to potential differentiation stimuli; (b) distinguishing MSCs from TM cells; (c) distinguishing subtypes of TM cells; and (d) distinguishing TM cells from those in neighbouring tissue. At the protein level, dexamethasone induction of myocilin was a robust discriminating factor between MSCs and TM cells and was complemented by other protein markers. Finally, we show that contractility and phagocytosis differ between MSCs and TM cells. These methods are recommended for use in future differentiation studies to fully define if a functional TM-like phenotype is being achieved.
Keyphrases
- induced apoptosis
- cell cycle arrest
- stem cells
- mesenchymal stem cells
- cell therapy
- oxidative stress
- endoplasmic reticulum stress
- type diabetes
- single cell
- flow cytometry
- transcription factor
- metabolic syndrome
- low dose
- newly diagnosed
- high resolution
- skeletal muscle
- prognostic factors
- body composition
- pi k akt
- cell proliferation
- protein protein
- risk assessment
- optic nerve
- current status
- human health
- smooth muscle