Human conjunctival epithelial cell expansion on mitotically inactivated 3T3 feeder cells was performed using methods described previously for limbal epithelial cell expansion (Collin et al., 2021a (link)). Cell colonies with typical epithelial morphology started to appear after 4–7 days and were cultured until they became subconfluent. Following this, 3T3 feeder cells were detached and removed using 0.02% EDTA (Lonza, Switzerland), and subconfluent primary cultures were dissociated with 0.5% trypsin-EDTA (Santa Cruz, CA, USA) to single-cell suspension and passaged at a density of 6 × 103 cells/cm2.
Trypsin edta
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Trypsin-EDTA is a solution used to dissociate and detach adherent cells in cell culture applications. It contains the enzyme trypsin and the chelating agent EDTA, which work together to break down the cell-cell and cell-substrate attachments, allowing cells to be suspended for further processing or subculturing.
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Lab products found in correlation
Ethylene diamine tetraacetic acid (edta), by Lonza (1 mentions)
Mmp 9, by Santa Cruz Biotechnology (1 mentions)
Penicillin streptomycin, by Santa Cruz Biotechnology (1 mentions)
Timp 2, by Santa Cruz Biotechnology (1 mentions)
Timp 1, by Santa Cruz Biotechnology (1 mentions)
Matrigel, by BD (1 mentions)
2 protocols using trypsin edta
1
Expansion of Human Conjunctival Epithelial Cells
2
Hedgehog Signaling Pathway Analysis
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Dulbecco's modified Eagle medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin, trypsin-EDTA, antibodies for SHH (Human, pAd), PTCH (Human, pAd), SMO (Human, pAd), Gli1 (Human, pAd), MMP2 (Human, pAd), MMP9 (Human, pAd), TIMP1 (Human, pAd), and TIMP2 (Human, pAd) were obtained from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). All antibodies were diluted 1:100. Matrigel was purchased from BD Biosciences (Bedford, MA). Eight-micrometer pore size transwell inserts were purchased from Corning Corporation (Corning, NY). All other chemicals, unless otherwise stated, were obtained from Sigma Chemicals (St Louis, MO).
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