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Survival of human embryonic stem cells implanted in the guinea pig auditory epithelium

View Article: PubMed Central - PubMed

ABSTRACT

Hair cells in the mature cochlea cannot spontaneously regenerate. One potential approach for restoring hair cells is stem cell therapy. However, when cells are transplanted into scala media (SM) of the cochlea, they promptly die due to the high potassium concentration. We previously described a method for conditioning the SM to make it more hospitable to implanted cells and showed that HeLa cells could survive for up to a week using this method. Here, we evaluated the survival of human embryonic stem cells (hESC) constitutively expressing GFP (H9 Cre-LoxP) in deaf guinea pig cochleae that were pre-conditioned to reduce potassium levels. GFP-positive cells could be detected in the cochlea for at least 7 days after the injection. The cells appeared spherical or irregularly shaped, and some were aggregated. Flushing SM with sodium caprate prior to transplantation resulted in a lower proportion of stem cells expressing the pluripotency marker Oct3/4 and increased cell survival. The data demonstrate that conditioning procedures aimed at transiently reducing the concentration of potassium in the SM facilitate survival of hESCs for at least one week. During this time window, additional procedures can be applied to initiate the differentiation of the implanted hESCs into new hair cells.

No MeSH data available.


Distribution and density of transplanted hESCs at 7 days.All morphologies were widely distributed at 1 day (a), but had narrower distributions at 7 days (b). Total number of GFP-positive cells declined between 1 day and 7 days (c).
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f7: Distribution and density of transplanted hESCs at 7 days.All morphologies were widely distributed at 1 day (a), but had narrower distributions at 7 days (b). Total number of GFP-positive cells declined between 1 day and 7 days (c).

Mentions: To test whether sodium caprate enhanced longer-term hESC survival, we extended the post-inoculation survival time to 7 days and assessed the distribution and density of the transplanted hESCs. All stem cell morphologies were widely distributed at 1 day (Fig. 7a), but had narrower distributions at 7 days (Fig. 7b). At the 7-day time point, GFP-positive cells were mostly observed near the cochleostomy site (5 mm away from apical end). The total number of GFP-positive cells was lower at 7 days than at 1 day (Fig. 7c), but the difference was not statistically significant (p = 0.489).


Survival of human embryonic stem cells implanted in the guinea pig auditory epithelium
Distribution and density of transplanted hESCs at 7 days.All morphologies were widely distributed at 1 day (a), but had narrower distributions at 7 days (b). Total number of GFP-positive cells declined between 1 day and 7 days (c).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5384248&req=5

f7: Distribution and density of transplanted hESCs at 7 days.All morphologies were widely distributed at 1 day (a), but had narrower distributions at 7 days (b). Total number of GFP-positive cells declined between 1 day and 7 days (c).
Mentions: To test whether sodium caprate enhanced longer-term hESC survival, we extended the post-inoculation survival time to 7 days and assessed the distribution and density of the transplanted hESCs. All stem cell morphologies were widely distributed at 1 day (Fig. 7a), but had narrower distributions at 7 days (Fig. 7b). At the 7-day time point, GFP-positive cells were mostly observed near the cochleostomy site (5 mm away from apical end). The total number of GFP-positive cells was lower at 7 days than at 1 day (Fig. 7c), but the difference was not statistically significant (p = 0.489).

View Article: PubMed Central - PubMed

ABSTRACT

Hair cells in the mature cochlea cannot spontaneously regenerate. One potential approach for restoring hair cells is stem cell therapy. However, when cells are transplanted into scala media (SM) of the cochlea, they promptly die due to the high potassium concentration. We previously described a method for conditioning the SM to make it more hospitable to implanted cells and showed that HeLa cells could survive for up to a week using this method. Here, we evaluated the survival of human embryonic stem cells (hESC) constitutively expressing GFP (H9 Cre-LoxP) in deaf guinea pig cochleae that were pre-conditioned to reduce potassium levels. GFP-positive cells could be detected in the cochlea for at least 7 days after the injection. The cells appeared spherical or irregularly shaped, and some were aggregated. Flushing SM with sodium caprate prior to transplantation resulted in a lower proportion of stem cells expressing the pluripotency marker Oct3/4 and increased cell survival. The data demonstrate that conditioning procedures aimed at transiently reducing the concentration of potassium in the SM facilitate survival of hESCs for at least one week. During this time window, additional procedures can be applied to initiate the differentiation of the implanted hESCs into new hair cells.

No MeSH data available.