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Lipopolysaccharide preconditioning of adipose-derived stem cells improves liver-regenerating activity of the secretome.

Lee SC, Jeong HJ, Lee SK, Kim SJ - Stem Cell Res Ther (2015)

Bottom Line: LPS-CM significantly promoted thioacetamide-damaged AML12 cell viability compared with CM-incubated cells and the control cells (77%, 69%, and 65% P<0.05).In the in vivo experiment, LPS-CM infusion into the partially hepatectomized mice significantly reduced serum IL-6 and TNF-α levels compared with the other groups (P<0.05) on days 1 and 2 after partial hepatectomy.Our results suggest that LPS preconditioning effectively stimulates ASCs to produce the secretome beneficial to hepatic regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Daejeon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Daeheung-dong 520-2, Joong-gu, Daejeon, Republic of Korea. zambo9@catholic.ac.kr.

ABSTRACT

Introduction: Growing recognition of paracrine mechanisms in stem cell plasticity has resulted in considerable interest in stem cell-derived secretome. The aim of this study was to investigate the effects of lipopolysaccharide (LPS) preconditioning on the composition and hepatic regenerative activity of adipose-derived stem cell (ASC) secretome.

Methods: Conditioned medium (CM) and LPS-CM were obtained after culturing human ASCs without or with low-dose LPS (0.5 ng/mL) for 24 hours. Untreated and thioacetamide-treated mouse AML12 hepatocytes were incubated for 24 hours with the control medium, LPS (0.5 ng/mL), CM, and LPS-CM and then cell viabilities were compared. CM and LPS-CM were also intravenously administered to partially hepatectomized mice, and their effects on liver regeneration were assessed by using liver weight measurements, immunohistochemistry, and Western blotting.

Results: In the in vitro experiments, LPS preconditioning of ASCs enhanced the mRNA expression levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), hepatocyte growth factor, and vascular endothelial growth factor, which evoke inflammatory response or liver regeneration. LPS-CM significantly promoted thioacetamide-damaged AML12 cell viability compared with CM-incubated cells and the control cells (77%, 69%, and 65% P<0.05). In the in vivo experiment, LPS-CM infusion into the partially hepatectomized mice significantly reduced serum IL-6 and TNF-α levels compared with the other groups (P<0.05) on days 1 and 2 after partial hepatectomy. Moreover, LPS-CM infusion enhanced liver regeneration (based on the liver weight changes at day 7 after partial hepatectomy, 3.73% versus 3.22% in the CM group; P<0.05) and significantly reduced the elevated serum levels of aspartate transaminase and alanine transaminase (at day 1, P<0.05).

Conclusions: Our results suggest that LPS preconditioning effectively stimulates ASCs to produce the secretome beneficial to hepatic regeneration. Thus, optimizing ASC secretome profile by LPS preconditioning could be a promising approach to treat liver diseases by using stem cells.

No MeSH data available.


Related in: MedlinePlus

Effect of conditioned medium (CM) and lipopolysaccharide (LPS)-CM on serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) in partially hepatectomized mice. Mice were intravenously injected with low-glucose Dulbecco’s modified Eagle’s medium (control), LPS (0.5 ng/mL), CM, or LPS-CM 1 hour after partial hepatectomy. At day 1 after partial hepatectomy, the LPS-CM-treated mice had significantly lower AST (A) and ALT (B) levels compared with the other groups (P <0.05). *P <0.05.
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Fig5: Effect of conditioned medium (CM) and lipopolysaccharide (LPS)-CM on serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) in partially hepatectomized mice. Mice were intravenously injected with low-glucose Dulbecco’s modified Eagle’s medium (control), LPS (0.5 ng/mL), CM, or LPS-CM 1 hour after partial hepatectomy. At day 1 after partial hepatectomy, the LPS-CM-treated mice had significantly lower AST (A) and ALT (B) levels compared with the other groups (P <0.05). *P <0.05.

Mentions: Antigen Ki67 is a nuclear protein associated with the transcription of ribosomal RNA and therefore is expressed exclusively in proliferating cells [40]. Figure 4A shows the representative sections of Ki67-labeled hepatocytes on day 1 after PH. The number of Ki67-positive cells peaked on day 2 and decreased thereafter in all the groups (Figure 5B). The LPS-CM mice demonstrated a significantly higher number of Ki67-positive cells in the liver compared with the control, LPS, and CM mice on day 1, 2, and 3 after PH (P <0.05). However, on day 7, the difference in the number of Ki67-positive cells between the CM and LPS-CM groups became insignificant.Figure 5


Lipopolysaccharide preconditioning of adipose-derived stem cells improves liver-regenerating activity of the secretome.

Lee SC, Jeong HJ, Lee SK, Kim SJ - Stem Cell Res Ther (2015)

Effect of conditioned medium (CM) and lipopolysaccharide (LPS)-CM on serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) in partially hepatectomized mice. Mice were intravenously injected with low-glucose Dulbecco’s modified Eagle’s medium (control), LPS (0.5 ng/mL), CM, or LPS-CM 1 hour after partial hepatectomy. At day 1 after partial hepatectomy, the LPS-CM-treated mice had significantly lower AST (A) and ALT (B) levels compared with the other groups (P <0.05). *P <0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4416308&req=5

Fig5: Effect of conditioned medium (CM) and lipopolysaccharide (LPS)-CM on serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) in partially hepatectomized mice. Mice were intravenously injected with low-glucose Dulbecco’s modified Eagle’s medium (control), LPS (0.5 ng/mL), CM, or LPS-CM 1 hour after partial hepatectomy. At day 1 after partial hepatectomy, the LPS-CM-treated mice had significantly lower AST (A) and ALT (B) levels compared with the other groups (P <0.05). *P <0.05.
Mentions: Antigen Ki67 is a nuclear protein associated with the transcription of ribosomal RNA and therefore is expressed exclusively in proliferating cells [40]. Figure 4A shows the representative sections of Ki67-labeled hepatocytes on day 1 after PH. The number of Ki67-positive cells peaked on day 2 and decreased thereafter in all the groups (Figure 5B). The LPS-CM mice demonstrated a significantly higher number of Ki67-positive cells in the liver compared with the control, LPS, and CM mice on day 1, 2, and 3 after PH (P <0.05). However, on day 7, the difference in the number of Ki67-positive cells between the CM and LPS-CM groups became insignificant.Figure 5

Bottom Line: LPS-CM significantly promoted thioacetamide-damaged AML12 cell viability compared with CM-incubated cells and the control cells (77%, 69%, and 65% P<0.05).In the in vivo experiment, LPS-CM infusion into the partially hepatectomized mice significantly reduced serum IL-6 and TNF-α levels compared with the other groups (P<0.05) on days 1 and 2 after partial hepatectomy.Our results suggest that LPS preconditioning effectively stimulates ASCs to produce the secretome beneficial to hepatic regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Daejeon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Daeheung-dong 520-2, Joong-gu, Daejeon, Republic of Korea. zambo9@catholic.ac.kr.

ABSTRACT

Introduction: Growing recognition of paracrine mechanisms in stem cell plasticity has resulted in considerable interest in stem cell-derived secretome. The aim of this study was to investigate the effects of lipopolysaccharide (LPS) preconditioning on the composition and hepatic regenerative activity of adipose-derived stem cell (ASC) secretome.

Methods: Conditioned medium (CM) and LPS-CM were obtained after culturing human ASCs without or with low-dose LPS (0.5 ng/mL) for 24 hours. Untreated and thioacetamide-treated mouse AML12 hepatocytes were incubated for 24 hours with the control medium, LPS (0.5 ng/mL), CM, and LPS-CM and then cell viabilities were compared. CM and LPS-CM were also intravenously administered to partially hepatectomized mice, and their effects on liver regeneration were assessed by using liver weight measurements, immunohistochemistry, and Western blotting.

Results: In the in vitro experiments, LPS preconditioning of ASCs enhanced the mRNA expression levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), hepatocyte growth factor, and vascular endothelial growth factor, which evoke inflammatory response or liver regeneration. LPS-CM significantly promoted thioacetamide-damaged AML12 cell viability compared with CM-incubated cells and the control cells (77%, 69%, and 65% P<0.05). In the in vivo experiment, LPS-CM infusion into the partially hepatectomized mice significantly reduced serum IL-6 and TNF-α levels compared with the other groups (P<0.05) on days 1 and 2 after partial hepatectomy. Moreover, LPS-CM infusion enhanced liver regeneration (based on the liver weight changes at day 7 after partial hepatectomy, 3.73% versus 3.22% in the CM group; P<0.05) and significantly reduced the elevated serum levels of aspartate transaminase and alanine transaminase (at day 1, P<0.05).

Conclusions: Our results suggest that LPS preconditioning effectively stimulates ASCs to produce the secretome beneficial to hepatic regeneration. Thus, optimizing ASC secretome profile by LPS preconditioning could be a promising approach to treat liver diseases by using stem cells.

No MeSH data available.


Related in: MedlinePlus