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Mesenchymal stem cells administered in the early phase of tumorigenesis inhibit colorectal tumor development in rats.

Katsuno T, Ochi M, Tominaga K, Tanaka F, Sogawa M, Tanigawa T, Yamagami H, Shiba M, Watanabe K, Watanabe T, Fujiwara Y, Arakawa T - J Clin Biochem Nutr (2013)

Bottom Line: To investigate the differences between the effects of mesenchymal stem cells (MSCs) administered in the early and late phases of tumorigenesis, MSCs were isolated from bone marrow and colorectal tumors were produced by exposing 7-week-old F344 rats to 1,2-dimethylhydrazine and dextran sulfate sodium.Administered MSCs in the early phase reduced ACF number on days 14 and 35 (1.9 vs 4.1 and 3.7 vs 7.3; p<0.01).Rectal TGF-β1 increased 1.3 fold on day 3, and MSC-conditioned medium containing TGF-β1 abundantly inhibited ACL15 cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka 545-8585, Japan.

ABSTRACT
To investigate the differences between the effects of mesenchymal stem cells (MSCs) administered in the early and late phases of tumorigenesis, MSCs were isolated from bone marrow and colorectal tumors were produced by exposing 7-week-old F344 rats to 1,2-dimethylhydrazine and dextran sulfate sodium. We evaluated tumor number and volume (week 25), MSC localization, number of aberrant crypt foci (ACF), transforming growth factor (TGF)-β1 protein levels in the rectum after administration of MSCs (week 5 or 15), and the effects of MSC-conditioned medium on ACL15 cell proliferation. Administered MSCs labeled with PKH26 were observed in the rectum. Administered MSCs in the early phase (week 5) before tumor occurrence (week 12) significantly decreased tumor number and volume (1.5 vs 4 and 21 mm(3) vs 170 mm(3); p<0.01), but not administered MSCs in the late phase (week 15). Administered MSCs in the early phase reduced ACF number on days 14 and 35 (1.9 vs 4.1 and 3.7 vs 7.3; p<0.01). Rectal TGF-β1 increased 1.3 fold on day 3, and MSC-conditioned medium containing TGF-β1 abundantly inhibited ACL15 cell proliferation. MSCs administered in the early phase but not late phase inhibited colorectal tumor development in a rat model.

No MeSH data available.


Related in: MedlinePlus

Characterization of MSCs. Characterization of MSCs was identified by flow cytometry using antibodies against CD29, CD34, CD44, CD45, and CD90. MSC cells were positive for CD29, CD44, and CD90, but negative for CD34 and CD45.
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Figure 1: Characterization of MSCs. Characterization of MSCs was identified by flow cytometry using antibodies against CD29, CD34, CD44, CD45, and CD90. MSC cells were positive for CD29, CD44, and CD90, but negative for CD34 and CD45.

Mentions: MSCs were harvested and cultured as described previously.(7) Briefly, 7-week-old male F344 rats were sacrificed by cervical dislocation, and bone marrow was obtained from the tibia and femur of rats. MSCs were cultured in low-glucose Dulbecco’s modified Eagle’s medium (DMEM: Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Carlsbad, CA). All cells were incubated at 37°C in a 5% CO2 humidified cell culture incubator. Non-adherent cells were removed by changing the medium at 72 h every 4 days. MSCs morphologically appeared to be a homogenous population of spindle-shaped cells such as fibroblasts. These cells were used in subsequent experiments after the fifth passage. To evaluate the lineage, MSCs were washed and fixed and permeabilized by a fixation buffer and permeabilization buffer, and then stained with anti-CD29 (GenWay Biotech Inc, San Diego, CA), anti-CD44 (Acris Antibodies, Hiddenhausen, Germany), anti-CD90 (AbD Serotec, Oxford, UK), anti-CD34 (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-CD45 (Acris Antibodies) for 30 min at 4°C.(22) The cells were washed, and a LSR II flow cytometer (BD Biosciences, San Jose, CA) was used to confirm that these cells expressed CD29, CD44, and CD90 and not CD34 or CD45 (Fig. 1).


Mesenchymal stem cells administered in the early phase of tumorigenesis inhibit colorectal tumor development in rats.

Katsuno T, Ochi M, Tominaga K, Tanaka F, Sogawa M, Tanigawa T, Yamagami H, Shiba M, Watanabe K, Watanabe T, Fujiwara Y, Arakawa T - J Clin Biochem Nutr (2013)

Characterization of MSCs. Characterization of MSCs was identified by flow cytometry using antibodies against CD29, CD34, CD44, CD45, and CD90. MSC cells were positive for CD29, CD44, and CD90, but negative for CD34 and CD45.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818273&req=5

Figure 1: Characterization of MSCs. Characterization of MSCs was identified by flow cytometry using antibodies against CD29, CD34, CD44, CD45, and CD90. MSC cells were positive for CD29, CD44, and CD90, but negative for CD34 and CD45.
Mentions: MSCs were harvested and cultured as described previously.(7) Briefly, 7-week-old male F344 rats were sacrificed by cervical dislocation, and bone marrow was obtained from the tibia and femur of rats. MSCs were cultured in low-glucose Dulbecco’s modified Eagle’s medium (DMEM: Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Carlsbad, CA). All cells were incubated at 37°C in a 5% CO2 humidified cell culture incubator. Non-adherent cells were removed by changing the medium at 72 h every 4 days. MSCs morphologically appeared to be a homogenous population of spindle-shaped cells such as fibroblasts. These cells were used in subsequent experiments after the fifth passage. To evaluate the lineage, MSCs were washed and fixed and permeabilized by a fixation buffer and permeabilization buffer, and then stained with anti-CD29 (GenWay Biotech Inc, San Diego, CA), anti-CD44 (Acris Antibodies, Hiddenhausen, Germany), anti-CD90 (AbD Serotec, Oxford, UK), anti-CD34 (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-CD45 (Acris Antibodies) for 30 min at 4°C.(22) The cells were washed, and a LSR II flow cytometer (BD Biosciences, San Jose, CA) was used to confirm that these cells expressed CD29, CD44, and CD90 and not CD34 or CD45 (Fig. 1).

Bottom Line: To investigate the differences between the effects of mesenchymal stem cells (MSCs) administered in the early and late phases of tumorigenesis, MSCs were isolated from bone marrow and colorectal tumors were produced by exposing 7-week-old F344 rats to 1,2-dimethylhydrazine and dextran sulfate sodium.Administered MSCs in the early phase reduced ACF number on days 14 and 35 (1.9 vs 4.1 and 3.7 vs 7.3; p<0.01).Rectal TGF-β1 increased 1.3 fold on day 3, and MSC-conditioned medium containing TGF-β1 abundantly inhibited ACL15 cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka 545-8585, Japan.

ABSTRACT
To investigate the differences between the effects of mesenchymal stem cells (MSCs) administered in the early and late phases of tumorigenesis, MSCs were isolated from bone marrow and colorectal tumors were produced by exposing 7-week-old F344 rats to 1,2-dimethylhydrazine and dextran sulfate sodium. We evaluated tumor number and volume (week 25), MSC localization, number of aberrant crypt foci (ACF), transforming growth factor (TGF)-β1 protein levels in the rectum after administration of MSCs (week 5 or 15), and the effects of MSC-conditioned medium on ACL15 cell proliferation. Administered MSCs labeled with PKH26 were observed in the rectum. Administered MSCs in the early phase (week 5) before tumor occurrence (week 12) significantly decreased tumor number and volume (1.5 vs 4 and 21 mm(3) vs 170 mm(3); p<0.01), but not administered MSCs in the late phase (week 15). Administered MSCs in the early phase reduced ACF number on days 14 and 35 (1.9 vs 4.1 and 3.7 vs 7.3; p<0.01). Rectal TGF-β1 increased 1.3 fold on day 3, and MSC-conditioned medium containing TGF-β1 abundantly inhibited ACL15 cell proliferation. MSCs administered in the early phase but not late phase inhibited colorectal tumor development in a rat model.

No MeSH data available.


Related in: MedlinePlus