Summary
A study of porcine iPS cells in an SLA-matched setting is a possible robust model for transplantation of human iPS cells in an HLA-matched setting.

Induced pluripotent stem cells generated from adult mature cells have changed the way we look at the mechanisms of disease and develop therapies. One area of promising potential is using iPS cells for autologous stem cell therapies. However, autologous derivation and clinical efficacy testing of iPS cells is a considerable effort involving isolating functionally active cells.

One approach was used by Mizukami, Abe, Shibata, et al1 from Jichi Medical University, Tsukuba Primate Research Center, and the Japan Science and Technology Agency, assessing immune responses to iPS cells in an MHC-matched setting through use of inbred SLA-defined miniature swine.

Their study tried to determine if transplantation of porcine iPS cells in an SLA-matched setting might provide a robust model for transplantation of human iPS cells in an HLA-matched setting. They used various approaches including immunohistochemistry, mixed lymphocyte reaction, in vitro phagocytosis assays, humoral immune responses, and NK cell mediated cytotoxicity assays.

Feeder cells present in donor cells
Figure 1. Feeder cells present in donor cells. Flow cytometric analysis showed that feeder cells were present in donor cells. EGFP-labeled C1 iPS cells were used to distinguish from mouse feeder cells (STO). C1 iPS cells were harvested by trypsinization and then incubated on gelatin-coated dishes for 15 min to remove feeder cells. 27.4% of the harvested cells were feeder cells. Source: MHC-Matched Induced Pluripotent Stem Cells Can Attenuate Cellular and Humoral Immune Responses but Are Still Susceptible to Innate Immunity in Pigs by Mizukami et al is licensed by CC BY 4.0.

 

To perform the NK mediated cytotoxicity assay, the Sony SH800 cell sorter was used to isolate CD3-negative and CD16-positive cells from donor pigs. This assay is highly sensitive, and the SH800 cell sorter successfully sorted a highly pure and functionally active NK cell population. Figure 1 shows the results of the flow cytometrics analysis that feeder cells were present in donor cells.

 

➤ Learn more about how other labs use cell sorting for developmental biology research

 


References

Mizukami, Abe, Shibata, et al. MHC-matched induced pluripotent stem cells can attenuate cellular and humoral immune responses but are still susceptible to innate immunity in pigs. PLOS One June 2014 | Volume 9 | Issue 6. PubMed