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Updated: Jul 17, 2026
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Cryopreservation of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells at the Optimal Stage
Published on: November 3, 2023
Effective cryopreservation of human embryonic stem cells by programmed freezing
Peng-Fei Yang1, Tse-Chao Hua, H-C Tsung
1Shanghai University of Science and Technology, No.516,Jun Gong Road,200093,Shanghai, China.
Summary
Cryopreservation of human embryonic stem cells (ES cells) is challenging. A novel method using a programmable cooler and specific cryoprotective agents achieved an 81.8% survival rate for ES cells, maintaining their pluripotency.
Area of Science:
- Stem Cell Biology
- Cryobiology
- Biotechnology
Background:
- Cryopreservation of human embryonic stem cells (ES cells) remains a significant challenge in regenerative medicine and research.
- Maintaining the viability and pluripotency of ES cells post-cryopreservation is crucial for their therapeutic and research applications.
Purpose of the Study:
- To develop and optimize a cryopreservation protocol for human ES cells.
- To evaluate the survival rate and pluripotency of cryopreserved human ES cells.
Main Methods:
- Utilized a computer-controlled programmable cooler for controlled cooling.
- Investigated the effects of cooling rate, seeding temperature, and cryoprotective agent composition (Me2SO+FBS+DMEM in a 1:3:6 ratio).
- Plunged samples into liquid nitrogen immediately after reaching the terminative temperature (-35°C from 0°C at 0.5°C/min, seeding at -10°C).
Main Results:
- Achieved a high survival rate of 81.8% for cryopreserved human ES cells.
- Cryopreserved cells retained pluripotency after prolonged cultivation.
- Normal karyotype and positive alkaline phosphatase staining confirmed cell viability and stem cell characteristics.
Conclusions:
- The optimized cryopreservation protocol significantly improves human ES cell survival rates.
- Cryopreserved human ES cells maintain their essential properties, enabling long-term storage and future use.
- This method offers a reliable approach for preserving valuable human ES cell lines.
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00:58Embryonic Stem Cells
Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.
00:57Embryonic Stem Cells
Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
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01:28Cryo-electron Microscopy
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