Human feeder layer system derived from umbilical cord stromal cells for human embryonic stem cells

Abstract

Objective: To evaluate the potential of human umbilical cord-derived stromal cells (hUCSCs) as a human feeder for human embryonic stem cells (ESCs).

Design: Prospective study.

Setting: Laboratory of Molecular Genetics and Stem Cell Differentiation, Dental Research Institute, School of Dentistry, Seoul National University.

Intervention(s): The hUCSCs were established, and human ESCs were cultured on established hUCSCs without serum.

Main outcome measure(s): Cell-surface markers, karyotyping, and teratoma formation.

Result(s): Primary cultures of hUCSCs from individual umbilical cords were maintained by an established protocol. Human ESCs on hUCSC layers were successfully maintained in serum-free culture medium past passage 30. Compared with hESCs on mouse feeder cells, the hESCs on hUCSCs showed similar levels of pluripotency-related cell-surface markers, self-renewal capacity, and teratoma formation in immune-deficient mice. These ESCs cultured on hUCSCs had a normal karyotype, even after long-term culture.

Conclusion(s): The hUCSCs supported self-renewal of hESCs in serum-free conditions. This culture system has the potential to facilitate the development of clinical-grade hESCs for regenerative medicine.