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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

Experimental protocols, macroscopic images of a rat colorectal tumor model and localization of exogenously administered MSCs. (A) Experimental protocol to confirm the period of formation and location of tumor formation. (B) Experimental protocol to confirm the effects of administering MSCs on colorectal tumors between early and late phases of tumorigenesis. ↑, subcutaneous injection of 1,2-dimethylhydrazine (DMH; 40 mg/kg); ■, duration of the free drinking of water containing 1.0% dextran sulfate sodium (DSS) during the second week; ▲, the intravenous injection of MSCs (4 × 106 cells/body), and †, sacrificed rats. (C) Tumors at week 25. (a) normal group; (b) control group; (c) MSCs group (administered at week 5); and (d) MSCs group (administered at week 15). (D) Exogenously administered MSCs with PKH26 (red) was localized the lamina propria, particularly near the bottom of the crypt, on day 3 after the administration of MSCs (blue: DAPI staining) (40× magnification).
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Figure 2: Experimental protocols, macroscopic images of a rat colorectal tumor model and localization of exogenously administered MSCs. (A) Experimental protocol to confirm the period of formation and location of tumor formation. (B) Experimental protocol to confirm the effects of administering MSCs on colorectal tumors between early and late phases of tumorigenesis. ↑, subcutaneous injection of 1,2-dimethylhydrazine (DMH; 40 mg/kg); ■, duration of the free drinking of water containing 1.0% dextran sulfate sodium (DSS) during the second week; ▲, the intravenous injection of MSCs (4 × 106 cells/body), and †, sacrificed rats. (C) Tumors at week 25. (a) normal group; (b) control group; (c) MSCs group (administered at week 5); and (d) MSCs group (administered at week 15). (D) Exogenously administered MSCs with PKH26 (red) was localized the lamina propria, particularly near the bottom of the crypt, on day 3 after the administration of MSCs (blue: DAPI staining) (40× magnification).

Mentions: The experimental protocol for the colorectal tumor model was performed using a previously described protocol shown in Fig. 2A.(23) Briefly, 1,2-dimethylhydrazine (DMH; 40 mg/kg body weight; Wako Pure Chem. Ind. Ltd., Osaka, Japan) was subcutaneously injected 3 times in the first week. Rats were then provided drinking water containing 1.0% dextran sulfate sodium (DSS) (Wako Pure Chem. Ind. Ltd.) every day during the second week. On week 5 or 15, the MSCs (4 × 106 cells/body with 1 ml saline) or saline alone (control) were administered to rats through the tail vein, and rats were sacrificed at week 25. The appropriate number of MSCs in the present study was determined according to the previous our report.(6) We evaluated tumor number and their total volume at week 25. Incidence of tumor was defined as a ratio per rat of which one or more nodules over than 1 mm in diameter was macroscopically recognized throughout the entire colon. The development of aberrant crypt foci (ACF), which are presumed to be the earliest identifiable preneoplastic lesions in the colorectal tumor model,(24–26) was also evaluated on days 0, 14, and 35 after the administration of MSCs on week 5 (Fig. 3A). Excised colonic tissues were cut open along the longitudinal median axis, fixed flat between 2 sheets of filter paper with 10% formalin (Wako Pure Chem. Ind. Ltd.), and then stained with 0.05% methylene blue for 6 min. The number of ACF per colon was counted under a light microscope at 40× magnification.


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)

Experimental protocols, macroscopic images of a rat colorectal tumor model and localization of exogenously administered MSCs. (A) Experimental protocol to confirm the period of formation and location of tumor formation. (B) Experimental protocol to confirm the effects of administering MSCs on colorectal tumors between early and late phases of tumorigenesis. ↑, subcutaneous injection of 1,2-dimethylhydrazine (DMH; 40 mg/kg); ■, duration of the free drinking of water containing 1.0% dextran sulfate sodium (DSS) during the second week; ▲, the intravenous injection of MSCs (4 × 106 cells/body), and †, sacrificed rats. (C) Tumors at week 25. (a) normal group; (b) control group; (c) MSCs group (administered at week 5); and (d) MSCs group (administered at week 15). (D) Exogenously administered MSCs with PKH26 (red) was localized the lamina propria, particularly near the bottom of the crypt, on day 3 after the administration of MSCs (blue: DAPI staining) (40× magnification).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3818273&req=5

Figure 2: Experimental protocols, macroscopic images of a rat colorectal tumor model and localization of exogenously administered MSCs. (A) Experimental protocol to confirm the period of formation and location of tumor formation. (B) Experimental protocol to confirm the effects of administering MSCs on colorectal tumors between early and late phases of tumorigenesis. ↑, subcutaneous injection of 1,2-dimethylhydrazine (DMH; 40 mg/kg); ■, duration of the free drinking of water containing 1.0% dextran sulfate sodium (DSS) during the second week; ▲, the intravenous injection of MSCs (4 × 106 cells/body), and †, sacrificed rats. (C) Tumors at week 25. (a) normal group; (b) control group; (c) MSCs group (administered at week 5); and (d) MSCs group (administered at week 15). (D) Exogenously administered MSCs with PKH26 (red) was localized the lamina propria, particularly near the bottom of the crypt, on day 3 after the administration of MSCs (blue: DAPI staining) (40× magnification).
Mentions: The experimental protocol for the colorectal tumor model was performed using a previously described protocol shown in Fig. 2A.(23) Briefly, 1,2-dimethylhydrazine (DMH; 40 mg/kg body weight; Wako Pure Chem. Ind. Ltd., Osaka, Japan) was subcutaneously injected 3 times in the first week. Rats were then provided drinking water containing 1.0% dextran sulfate sodium (DSS) (Wako Pure Chem. Ind. Ltd.) every day during the second week. On week 5 or 15, the MSCs (4 × 106 cells/body with 1 ml saline) or saline alone (control) were administered to rats through the tail vein, and rats were sacrificed at week 25. The appropriate number of MSCs in the present study was determined according to the previous our report.(6) We evaluated tumor number and their total volume at week 25. Incidence of tumor was defined as a ratio per rat of which one or more nodules over than 1 mm in diameter was macroscopically recognized throughout the entire colon. The development of aberrant crypt foci (ACF), which are presumed to be the earliest identifiable preneoplastic lesions in the colorectal tumor model,(24–26) was also evaluated on days 0, 14, and 35 after the administration of MSCs on week 5 (Fig. 3A). Excised colonic tissues were cut open along the longitudinal median axis, fixed flat between 2 sheets of filter paper with 10% formalin (Wako Pure Chem. Ind. Ltd.), and then stained with 0.05% methylene blue for 6 min. The number of ACF per colon was counted under a light microscope at 40× magnification.

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