Abstract:The epithelial cell rests of Malassez (ERM) are essential in preventing ankylosis between the alveolar bone and the tooth (dentoalveolar ankylosis). Despite extensive research, the mechanism by which ERM cells suppress ankylosis remains uncertain; perhaps its varied population is to reason. Therefore, in this study, eighteen unique clones of ERM (CRUDE) were isolated using the single-cell limiting dilution and designated as ERM 1–18. qRT-PCR, ELISA, and western blot analyses revealed that ERM-2 and -3 had the … Show more
“…Both morphometric parameters suggest a reduced surface attachment and less cell stretching of the respective cells. However, for other cell cultures it has been reported that cellular morphology and growing behavior have been changed more drastically upon clonal isolation, although it was not interpreted as a result of single-cell isolation [ 15 ]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Due to the statistical nature of obtaining a single cell after dilution, this clonal isolation method is comparatively inefficient and laborious [ 9 ]. However, due to its ease of use, it is still widely applied in the field of cell engineering [ 15 – 17 ].…”
Clonal isolation is an integral step of numerous workflows in genome editing and cell engineering. It comprises the isolation of a single progenitor cell from a defined cell line population with subsequent expansion to obtain a monoclonal cell population. This process is associated with transient loss of cell–cell contacts and absence of a multicellular microenvironment. Previous studies have revealed transcriptomic changes upon clonal isolation with cell line specific extent. Since transcriptome alterations are only partially reflected on the proteome level, we sought to investigate the impact of clonal isolation on the cellular proteome to a depth of > 6000 proteins in three established pancreatic cancer cell lines. We show that clonal isolation does have an impact on the cellular proteome, however, with cell line specific extent, affecting different biological processes, and also depending on the isolation method. We demonstrate a different impact of clonal isolation on mesenchymal- and epithelial-derived cell lines mainly affecting cell proliferation, metabolism, cell adhesion and cellular stress. The results bear relevance to the field of genomic editing and cell engineering and highlight the need to consider the impact of clonal isolation when interpreting data stemming from experiments that include this step.
“…Both morphometric parameters suggest a reduced surface attachment and less cell stretching of the respective cells. However, for other cell cultures it has been reported that cellular morphology and growing behavior have been changed more drastically upon clonal isolation, although it was not interpreted as a result of single-cell isolation [ 15 ]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Due to the statistical nature of obtaining a single cell after dilution, this clonal isolation method is comparatively inefficient and laborious [ 9 ]. However, due to its ease of use, it is still widely applied in the field of cell engineering [ 15 – 17 ].…”
Clonal isolation is an integral step of numerous workflows in genome editing and cell engineering. It comprises the isolation of a single progenitor cell from a defined cell line population with subsequent expansion to obtain a monoclonal cell population. This process is associated with transient loss of cell–cell contacts and absence of a multicellular microenvironment. Previous studies have revealed transcriptomic changes upon clonal isolation with cell line specific extent. Since transcriptome alterations are only partially reflected on the proteome level, we sought to investigate the impact of clonal isolation on the cellular proteome to a depth of > 6000 proteins in three established pancreatic cancer cell lines. We show that clonal isolation does have an impact on the cellular proteome, however, with cell line specific extent, affecting different biological processes, and also depending on the isolation method. We demonstrate a different impact of clonal isolation on mesenchymal- and epithelial-derived cell lines mainly affecting cell proliferation, metabolism, cell adhesion and cellular stress. The results bear relevance to the field of genomic editing and cell engineering and highlight the need to consider the impact of clonal isolation when interpreting data stemming from experiments that include this step.
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