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CD90- (Thy-1-) high selection enhances reprogramming capacity of murine adipose-derived mesenchymal stem cells.

Kawamoto K, Konno M, Nagano H, Nishikawa S, Tomimaru Y, Akita H, Hama N, Wada H, Kobayashi S, Eguchi H, Tanemura M, Ito T, Doki Y, Mori M, Ishii H - Dis. Markers (2013)

Bottom Line: CD90(Hi) ADSCs showed increased numbers of alkaline phosphatase-positive colonies compared with CD90(Lo) ADSCs.CD90(Hi) ADSCs had greater reprogramming capacity than CD90(Lo) ADSCs, suggesting that ADSCs have heterogeneous subpopulations.Thus, CD90(Hi) selection presents an effective strategy to isolate a highly suppressive subpopulation for stem cell-based tolerance induction therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.

ABSTRACT

Background: Mesenchymal stem cells (MSCs), including adipose tissue-derived mesenchymal stem cells (ADSC), are multipotent and can differentiate into various cell types possessing unique immunomodulatory features. Several clinical trials have demonstrated the safety and possible efficacy of MSCs in organ transplantation. Thus, stem cell therapy is promising for tolerance induction. In this study, we assessed the reprogramming capacity of murine ADSCs and found that CD90 (Thy-1), originally discovered as a thymocyte antigen, could be a useful marker for cell therapy.

Method: Murine ADSCs were isolated from B6 mice, sorted using a FACSAria cell sorter by selection of CD90(Hi) or CD90(Lo), and then transduced with four standard factors (4F; Oct4, Sox2, Klf4, and c-Myc).

Results: Unsorted, CD90(Hi)-sorted, and CD90(Lo)-sorted murine ADSCs were reprogrammed using standard 4F transduction. CD90(Hi) ADSCs showed increased numbers of alkaline phosphatase-positive colonies compared with CD90(Lo) ADSCs. The relative reprogramming efficiencies of unsorted, CD90(Hi)-sorted, and CD90(Lo)-sorted ADSCs were 100%, 116.5%, and 74.7%, respectively. CD90(Hi) cells were more responsive to reprogramming.

Conclusion: CD90(Hi) ADSCs had greater reprogramming capacity than CD90(Lo) ADSCs, suggesting that ADSCs have heterogeneous subpopulations. Thus, CD90(Hi) selection presents an effective strategy to isolate a highly suppressive subpopulation for stem cell-based tolerance induction therapy.

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Colony formation of the sorted cells. Phase (a and b) and alkaline phosphatase staining (c and d) of CD90Lo (a and c) and  CD90Hi (b and d) are shown.
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fig6: Colony formation of the sorted cells. Phase (a and b) and alkaline phosphatase staining (c and d) of CD90Lo (a and c) and  CD90Hi (b and d) are shown.

Mentions: To show the successful induction of the undifferentiated state, we employed immunocytochemistry of Oct4 and SSEA1. As shown in Figure 5, colonies were positive for Oct4 and SSEA1. Sorted and 4F-transduced ADSCs also exhibited colony formation. As shown in Figures 6(a) and 6(b), morphologically distinct colonies were visible in both CD90Hi and CD90Lo 4F-transduced ADSCs on posttransduction day 14. On posttransduction day 30, the colonies were stained with AP. There were AP+ colonies in both groups, although the CD90Hi ADSCs tended to form larger colonies than CD90Lo ADSCs (Figures 4(c) and 4(d)). As shown in Figure 7, CD90Hi ADSCs exhibited more AP+ colonies than  CD90Lo ADSCs. The reprogramming efficiencies of unsorted, CD90Hi-sorted, and CD90Lo-sorted ADSCs were 100%, 116.5%, and 74.7%, respectively.


CD90- (Thy-1-) high selection enhances reprogramming capacity of murine adipose-derived mesenchymal stem cells.

Kawamoto K, Konno M, Nagano H, Nishikawa S, Tomimaru Y, Akita H, Hama N, Wada H, Kobayashi S, Eguchi H, Tanemura M, Ito T, Doki Y, Mori M, Ishii H - Dis. Markers (2013)

Colony formation of the sorted cells. Phase (a and b) and alkaline phosphatase staining (c and d) of CD90Lo (a and c) and  CD90Hi (b and d) are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824355&req=5

fig6: Colony formation of the sorted cells. Phase (a and b) and alkaline phosphatase staining (c and d) of CD90Lo (a and c) and  CD90Hi (b and d) are shown.
Mentions: To show the successful induction of the undifferentiated state, we employed immunocytochemistry of Oct4 and SSEA1. As shown in Figure 5, colonies were positive for Oct4 and SSEA1. Sorted and 4F-transduced ADSCs also exhibited colony formation. As shown in Figures 6(a) and 6(b), morphologically distinct colonies were visible in both CD90Hi and CD90Lo 4F-transduced ADSCs on posttransduction day 14. On posttransduction day 30, the colonies were stained with AP. There were AP+ colonies in both groups, although the CD90Hi ADSCs tended to form larger colonies than CD90Lo ADSCs (Figures 4(c) and 4(d)). As shown in Figure 7, CD90Hi ADSCs exhibited more AP+ colonies than  CD90Lo ADSCs. The reprogramming efficiencies of unsorted, CD90Hi-sorted, and CD90Lo-sorted ADSCs were 100%, 116.5%, and 74.7%, respectively.

Bottom Line: CD90(Hi) ADSCs showed increased numbers of alkaline phosphatase-positive colonies compared with CD90(Lo) ADSCs.CD90(Hi) ADSCs had greater reprogramming capacity than CD90(Lo) ADSCs, suggesting that ADSCs have heterogeneous subpopulations.Thus, CD90(Hi) selection presents an effective strategy to isolate a highly suppressive subpopulation for stem cell-based tolerance induction therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.

ABSTRACT

Background: Mesenchymal stem cells (MSCs), including adipose tissue-derived mesenchymal stem cells (ADSC), are multipotent and can differentiate into various cell types possessing unique immunomodulatory features. Several clinical trials have demonstrated the safety and possible efficacy of MSCs in organ transplantation. Thus, stem cell therapy is promising for tolerance induction. In this study, we assessed the reprogramming capacity of murine ADSCs and found that CD90 (Thy-1), originally discovered as a thymocyte antigen, could be a useful marker for cell therapy.

Method: Murine ADSCs were isolated from B6 mice, sorted using a FACSAria cell sorter by selection of CD90(Hi) or CD90(Lo), and then transduced with four standard factors (4F; Oct4, Sox2, Klf4, and c-Myc).

Results: Unsorted, CD90(Hi)-sorted, and CD90(Lo)-sorted murine ADSCs were reprogrammed using standard 4F transduction. CD90(Hi) ADSCs showed increased numbers of alkaline phosphatase-positive colonies compared with CD90(Lo) ADSCs. The relative reprogramming efficiencies of unsorted, CD90(Hi)-sorted, and CD90(Lo)-sorted ADSCs were 100%, 116.5%, and 74.7%, respectively. CD90(Hi) cells were more responsive to reprogramming.

Conclusion: CD90(Hi) ADSCs had greater reprogramming capacity than CD90(Lo) ADSCs, suggesting that ADSCs have heterogeneous subpopulations. Thus, CD90(Hi) selection presents an effective strategy to isolate a highly suppressive subpopulation for stem cell-based tolerance induction therapy.

Show MeSH