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Comparing Two Intestinal Porcine Epithelial Cell Lines (IPECs): Morphological Differentiation, Function and Metabolism.

Nossol C, Barta-Böszörményi A, Kahlert S, Zuschratter W, Faber-Zuschratter H, Reinhardt N, Ponsuksili S, Wimmers K, Diesing AK, Rothkötter HJ - PLoS ONE (2015)

Bottom Line: On the other hand, "spliceosome", "ribosome", "RNA-degradation" and "tight junction" are significantly down-regulated pathways in IPEC-J2 in comparison to IPEC-1.These cells seem to be more active in their metabolism than IPEC-1 cells due to a significant higher ATP-content as well as a higher O2- and glucose-consumption.In addition, IPEC-J2 cells are a preferential tool for in vitro studies with the focus on metabolism.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Otto-von-Guericke University Magdeburg, 39120, Magdeburg, Germany.

ABSTRACT
The pig shows genetical and physiological resemblance to human, which predestines it as an experimental animal model especially for mucosal physiology. Therefore, the intestinal epithelial cell lines 1 and J2 (IPEC-1, IPEC-J2)--spontaneously immortalised cell lines from the porcine intestine--are important tools for studying intestinal function. A microarray (GeneChip Porcine Genome Array) was performed to compare the genome wide gene expression of IPECs. Different significantly up-regulated pathways were identified, like "lysosome", "pathways in cancer", "regulation of actin cytoskeleton" and "oxidative phosphorylation" in IPEC-J2 in comparison to IPEC-1. On the other hand, "spliceosome", "ribosome", "RNA-degradation" and "tight junction" are significantly down-regulated pathways in IPEC-J2 in comparison to IPEC-1. Examined pathways were followed up by functional analyses. ATP-, oxygen, glucose and lactate-measurement provide evidence for up-regulation of oxidative phosphorylation in IPEC-J2. These cells seem to be more active in their metabolism than IPEC-1 cells due to a significant higher ATP-content as well as a higher O2- and glucose-consumption. The down-regulated pathway "ribosome" was followed up by measurement of RNA- and protein content. In summary, IPEC-J2 is a morphologically and functionally more differentiated cell line in comparison to IPEC-1. In addition, IPEC-J2 cells are a preferential tool for in vitro studies with the focus on metabolism.

No MeSH data available.


Related in: MedlinePlus

Anchorage independent growth.Both cell lines were seeded in “Soft agar” (S, 0.33%) on “Feeder agar” (F, 0.5%) with (w) or without (wo) the application of EGF and ITS. Caco-2 cells were used as positive control. No anchorage independent growth was detected in IPEC-1 and IPEC-J2. On the other hand, Caco-2 showed an anchorage independent growth which did not depend on the additives EGF and ITS (bar = 20 μm). The results represent at least three independent experiments (n = 3).
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pone.0132323.g001: Anchorage independent growth.Both cell lines were seeded in “Soft agar” (S, 0.33%) on “Feeder agar” (F, 0.5%) with (w) or without (wo) the application of EGF and ITS. Caco-2 cells were used as positive control. No anchorage independent growth was detected in IPEC-1 and IPEC-J2. On the other hand, Caco-2 showed an anchorage independent growth which did not depend on the additives EGF and ITS (bar = 20 μm). The results represent at least three independent experiments (n = 3).

Mentions: IPEC-1 and IPEC-J2 are known to be non-transformed cell lines derived from normal intestinal cells. Typically, non-transformed epithelial cells do not grow in soft agar without solid cultivation surface. The anchorage independent growth was here used as a functional marker of tumorgenicity and compared with Caco-2 cells as a positive control. Caco-2 cells produced colonies within the agar independent of the additives of “feeder- and soft-agar”. In comparison to Caco-2, IPEC-1 and IPEC-J2 are non-infiltrating cells (Fig 1). After 3 weeks, only single cells and no colonies were detected. The supplementation of EGF and ITS (important growth factors for all three cell lines) to “feeder- and soft-agar” had no effects on the growth of IPECs.


Comparing Two Intestinal Porcine Epithelial Cell Lines (IPECs): Morphological Differentiation, Function and Metabolism.

Nossol C, Barta-Böszörményi A, Kahlert S, Zuschratter W, Faber-Zuschratter H, Reinhardt N, Ponsuksili S, Wimmers K, Diesing AK, Rothkötter HJ - PLoS ONE (2015)

Anchorage independent growth.Both cell lines were seeded in “Soft agar” (S, 0.33%) on “Feeder agar” (F, 0.5%) with (w) or without (wo) the application of EGF and ITS. Caco-2 cells were used as positive control. No anchorage independent growth was detected in IPEC-1 and IPEC-J2. On the other hand, Caco-2 showed an anchorage independent growth which did not depend on the additives EGF and ITS (bar = 20 μm). The results represent at least three independent experiments (n = 3).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493080&req=5

pone.0132323.g001: Anchorage independent growth.Both cell lines were seeded in “Soft agar” (S, 0.33%) on “Feeder agar” (F, 0.5%) with (w) or without (wo) the application of EGF and ITS. Caco-2 cells were used as positive control. No anchorage independent growth was detected in IPEC-1 and IPEC-J2. On the other hand, Caco-2 showed an anchorage independent growth which did not depend on the additives EGF and ITS (bar = 20 μm). The results represent at least three independent experiments (n = 3).
Mentions: IPEC-1 and IPEC-J2 are known to be non-transformed cell lines derived from normal intestinal cells. Typically, non-transformed epithelial cells do not grow in soft agar without solid cultivation surface. The anchorage independent growth was here used as a functional marker of tumorgenicity and compared with Caco-2 cells as a positive control. Caco-2 cells produced colonies within the agar independent of the additives of “feeder- and soft-agar”. In comparison to Caco-2, IPEC-1 and IPEC-J2 are non-infiltrating cells (Fig 1). After 3 weeks, only single cells and no colonies were detected. The supplementation of EGF and ITS (important growth factors for all three cell lines) to “feeder- and soft-agar” had no effects on the growth of IPECs.

Bottom Line: On the other hand, "spliceosome", "ribosome", "RNA-degradation" and "tight junction" are significantly down-regulated pathways in IPEC-J2 in comparison to IPEC-1.These cells seem to be more active in their metabolism than IPEC-1 cells due to a significant higher ATP-content as well as a higher O2- and glucose-consumption.In addition, IPEC-J2 cells are a preferential tool for in vitro studies with the focus on metabolism.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Otto-von-Guericke University Magdeburg, 39120, Magdeburg, Germany.

ABSTRACT
The pig shows genetical and physiological resemblance to human, which predestines it as an experimental animal model especially for mucosal physiology. Therefore, the intestinal epithelial cell lines 1 and J2 (IPEC-1, IPEC-J2)--spontaneously immortalised cell lines from the porcine intestine--are important tools for studying intestinal function. A microarray (GeneChip Porcine Genome Array) was performed to compare the genome wide gene expression of IPECs. Different significantly up-regulated pathways were identified, like "lysosome", "pathways in cancer", "regulation of actin cytoskeleton" and "oxidative phosphorylation" in IPEC-J2 in comparison to IPEC-1. On the other hand, "spliceosome", "ribosome", "RNA-degradation" and "tight junction" are significantly down-regulated pathways in IPEC-J2 in comparison to IPEC-1. Examined pathways were followed up by functional analyses. ATP-, oxygen, glucose and lactate-measurement provide evidence for up-regulation of oxidative phosphorylation in IPEC-J2. These cells seem to be more active in their metabolism than IPEC-1 cells due to a significant higher ATP-content as well as a higher O2- and glucose-consumption. The down-regulated pathway "ribosome" was followed up by measurement of RNA- and protein content. In summary, IPEC-J2 is a morphologically and functionally more differentiated cell line in comparison to IPEC-1. In addition, IPEC-J2 cells are a preferential tool for in vitro studies with the focus on metabolism.

No MeSH data available.


Related in: MedlinePlus