Abstract
Several studies have shown how different cell lines can influence the differentiation of stem cells through co-culture systems. The House Ear Institute-Organ of Corti 1 (HEI-OC1) is considered an important cell line for in vitro auditory research. However, it is unknown if HEI-OC1 cells can promote the differentiation of embryonic stem cells (ESCs). In this study, we investigated whether co-culture of ESCs with HEI-OC1 cells promotes differentiation. To this end, we developed a co-culture system of mouse ESCs with HEI-OC1 cells. Dissociated or embryonic bodies (EBs) of ESCs were introduced to a conditioned and inactivated confluent layer of HEI-OC1 cells for 14 days. The dissociated ESCs coalesced into an EB-like form that was smaller than the co-cultured EBs. Contact co-culture generated cells expressing several otic progenitor markers as well as hair cell specific markers. ESCs and EBs were also cultured in non-contact setup but using conditioned medium from HEI-OC1 cells, indicating that soluble factors alone could have a similar effect. The ESCs did not form into aggregates but were still Myo7a-positive, while the EBs degenerated. However, in the fully differentiated EBs, evidence to prove mature differentiation of inner ear hair cell was still rudimentary. Nevertheless, these results suggest that cellular interactions between ESCs and HEI-OC1 cells may both stimulate ESC differentiation.
Highlights
Several studies have shown how different cell lines can influence the differentiation of stem cells through co-culture systems
Initial passage of House Ear Institute-Organ of Corti 1 (HEI-OC1) cell culture at 37 °C and 5% CO2 cultured in embryonic stem cells (ESCs) medium had significantly low proliferative capacity relative to unconditioned cells kept at its typical medium (DMEM with 10% fetal bovine serum (FBS)) and incubated at 33 °C and 10% CO2
After five passages of conditioning, both cell proliferation and morphology were comparable to the unconditioned group (Fig. 1B) with a 98 ± % cell proliferation cultured in ESC media incubated at 37 °C and 5% C O2
Summary
Several studies have shown how different cell lines can influence the differentiation of stem cells through co-culture systems. The House Ear Institute-Organ of Corti 1 (HEI-OC1) is considered an important cell line for in vitro auditory research. It is unknown if HEI-OC1 cells can promote the differentiation of embryonic stem cells (ESCs). Aside from organotypic cell cultures of the inner ear, House Ear Institute-Organ of Corti 1 (HEI-OC1) cells bear the characteristics of cells of the sensory epithelium and are considered one of the most important cell lines available for in vitro auditory research. Pharmacological treatments related to hearing loss[10,11,12]
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