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Synovial fibroblasts derived from these cultures were routinely used in passages two or three, where these cells are known to conserve their phenotype [ 35].
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HUVECs were used in passages three to six.
Mutations were clustered according to their relative z-scores in passages one and two (Figure 4).
Cells in passages three to six were used for all experiments in this study.
All studies were performed using cultures in passage two.
Fibroblasts were cultivated in high-glucose DMEM Life Technologiess), Melanocytes in M2 (PromoCell) and assayed in passage four.
Treatment was performed on cells in passage six that were 70% confluent.
Notably the proportion of CD44+CD24low+ cells in passage four DT-22 was similar to passage 11 (representative data, Fig 1B).
Confluent cells in passage five were trypsinized and plated on eight-well slides (LabTek II Slide System, Nunc, Naperville, IL).
Treatment was performed on cells in passage six at 70% confluency, as routinely performed in our laboratory.
The cells showed no expression of CD45, CD20/CD79a, or HLA-DR, but were positive for CD73, CD90, and CD105 in passage three and six (Table 4).
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