P-MTAB-23641 - P-MTAB-23641

Primary human trabecular meshwork (HTM) HTM-72 and HTM-137 cell lines were generated respectively from the trabecular meshworks dissected from residual cornea rims of 29 and 39 years old donors (North Carolina Eye Bank) after surgical corneal transplants at the University of North Carolina Eye Clinic. The tissue was cut into small pieces, carefully attached to the bottom of the 2% gelatin-coated 35 mm dish, and covered slipped with a drop of MEM Richter’s Modification medium (IMEM, HyClone) supplemented with 20% fetal bovine serum (FBS), 50 ug/ml gentamicin (Invitrogen). Cells from these specimens were not treated with enzymes and were allowed to grow from the explant for a period of 4 weeks changing the media every other day; upon confluency, cells were harvested and stored in liquid nitrogen. When reconstituted, these primary nontransformed cells are grown in IMEM, 10% FBS, gentamicin and subsist for seven to eight passages. In this experiment study all cells were used at passage 4. These outflow pathway cultures comprise all cell types involved in maintaining resistance to flow. That includes cells from the three distinct regions of the trabecular meshwork plus cells lining the Schlemm’s canal. Because most of the cells in these cultures come from the trabecular meshwork, they are commonly referred to as “trabecular meshwork cells”. (Parameters: start time = 48, time unit = hours, min temperature = 37, temperature unit = C, media = MEM Richter’s Modification medium (IMEM))
Experiment E-MEXP-3427