TY - JOUR
T1 - Isolation and Characterization of primary human trabecular meshwork cells from segmental flow regions
T2 - New tools for understanding segmental flow
AU - Staverosky, Julia
AU - Dhamodaran, Kamesh
AU - Acott, Ted
AU - Raghunathan, Vijay Krishna
AU - Vranka, Janice
N1 - Funding Information:
The authors would like to thank their funding sources: NIH/NEI grants EY026048-01A1 (JAV, VKR), EY030238, EY008247, EY025721 (TSA), P30 EY010572, and by an unrestricted grant to the Casey Eye Institute from Research to Prevent Blindness, New York, NY. We would also like to thank the Lions VisionGift and SavingSight (Kansas City, MO) for procuring all human donor eyes used in this work. Most importantly, we would like to thank the families of the organ donors without whose consent these experiments would be impossible. We also would like to acknowledge the late John Bradley for his thoughtful comments and discussions over the years. His enthusiasm will be sorely missed.
Funding Information:
The authors would like to thank their funding sources: NIH / NEI grants EY026048-01A1 (JAV, VKR), EY030238 , EY008247 , EY025721 ( TSA ), P30 EY010572 , and by an unrestricted grant to the Casey Eye Institute from Research to Prevent Blindness , New York, NY. We would also like to thank the Lions VisionGift and SavingSight (Kansas City, MO) for procuring all human donor eyes used in this work. Most importantly, we would like to thank the families of the organ donors without whose consent these experiments would be impossible. We also would like to acknowledge the late John Bradley for his thoughtful comments and discussions over the years. His enthusiasm will be sorely missed.
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2020/8
Y1 - 2020/8
N2 - Segmental flow in the human trabecular meshwork is a well-documented phenomenon but in depth mechanistic investigations of high flow (HF) and low flow (LF) regions are restricted due to the small amount of tissue available from a single donor. To address this issue we have generated and characterized multiple paired HF and LF cell strains. Here paired HF and LF cell strains were generated from single donors. Cells were characterized for growth and proliferation, as well as gene and protein expression of potential segmental region markers. Cells isolated from HF and LF regions have similar growth and proliferation rates. Gene expression data reveals vascular cell adhesion protein 1 (VCAM1), thrombospondin 2 (THBS2), and tissue inhibitor of metalloproteinase 1 (TIMP1) are potential markers of LF cells in vitro. Protein expression of VCAM1, THBS2 and TIMP1 are complex and may reflect the dynamic nature of the TM. Initial protein expression levels of these genes is either similar between HF and LF cells (VCAM1, THBS2), or higher in HF compared to LF in some strains (TIMP1). However, after long term culture LF cells express higher levels of VCAM1, TIMP1 and THBS2 protein compared to HF cells. HF and LF cell strains are a powerful new tool that enable understanding segmental flow allowing for multiple experiments on the same genetic background.
AB - Segmental flow in the human trabecular meshwork is a well-documented phenomenon but in depth mechanistic investigations of high flow (HF) and low flow (LF) regions are restricted due to the small amount of tissue available from a single donor. To address this issue we have generated and characterized multiple paired HF and LF cell strains. Here paired HF and LF cell strains were generated from single donors. Cells were characterized for growth and proliferation, as well as gene and protein expression of potential segmental region markers. Cells isolated from HF and LF regions have similar growth and proliferation rates. Gene expression data reveals vascular cell adhesion protein 1 (VCAM1), thrombospondin 2 (THBS2), and tissue inhibitor of metalloproteinase 1 (TIMP1) are potential markers of LF cells in vitro. Protein expression of VCAM1, THBS2 and TIMP1 are complex and may reflect the dynamic nature of the TM. Initial protein expression levels of these genes is either similar between HF and LF cells (VCAM1, THBS2), or higher in HF compared to LF in some strains (TIMP1). However, after long term culture LF cells express higher levels of VCAM1, TIMP1 and THBS2 protein compared to HF cells. HF and LF cell strains are a powerful new tool that enable understanding segmental flow allowing for multiple experiments on the same genetic background.
KW - Anterior segment
KW - Aqueous humor outflow
KW - Glaucoma
KW - Segmental outflow
KW - Trabecular meshwork
UR - http://www.scopus.com/inward/record.url?scp=85085763552&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85085763552&partnerID=8YFLogxK
U2 - 10.1016/j.exer.2020.108046
DO - 10.1016/j.exer.2020.108046
M3 - Article
C2 - 32376472
AN - SCOPUS:85085763552
SN - 0014-4835
VL - 197
JO - Experimental Eye Research
JF - Experimental Eye Research
M1 - 108046
ER -