Cryopreservation of the Hair Follicle Maintains Pluripotency of Nestin-Expressing Hair Follicle-Associated Pluripotent Stem Cells

Tissue Engineering. Part C, Methods
Satoshi KajiuraYasuyuki Amoh

Abstract

Hair follicles contain nestin-expressing pluripotent stem cells, the origin of which is above the bulge area, below the sebaceous gland. We have termed these cells hair follicle-associated pluripotent (HAP) stem cells. In the present study, we established efficient cryopreservation methods of the hair follicle that maintained the pluripotency of HAP stem cells. We cryopreserved the whole hair follicle from green fluorescent protein transgenic mice by slow-rate cooling in TC-Protector medium and storage in liquid nitrogen. After thawing, the upper part of the hair follicle was isolated and cultured in Dulbecco's Modified Eagle's Medium (DMEM) with fetal bovine serum (FBS). After 4 weeks of culture, cells from the upper part of the hair follicle grew out. The growing cells were transferred to DMEM/F12 without FBS. After 1 week of culture, the growing cells formed hair spheres, each containing ∼1×10(2) HAP stem cells. The hair spheres contained cells that differentiated to neurons, glial cells, and other cell types. The thawed and cultured upper part of the hair follicle produced almost as many pluripotent hair spheres as fresh follicles. The hair spheres derived from slow-cooling cryopreserved hair follicles were as pluripotent a...Continue Reading

References

May 5, 1997·FEBS Letters·M OkabeY Nishimune
Jan 14, 2000·Nature Biotechnology·R M Hoffman
Aug 9, 2003·Proceedings of the National Academy of Sciences of the United States of America·Lingna LiRobert M Hoffman
Aug 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Yasuyuki AmohRobert M Hoffman
Apr 2, 2005·Proceedings of the National Academy of Sciences of the United States of America·Yasuyuki AmohRobert M Hoffman
Nov 30, 2005·Proceedings of the National Academy of Sciences of the United States of America·Yasuyuki AmohRobert M Hoffman
Jul 15, 2006·The Journal of Investigative Dermatology·Yasuyuki AmohRobert M Hoffman
Apr 6, 2011·Journal of Cellular Biochemistry·Aisada UchugonovaRobert M Hoffman
May 15, 2012·In Vitro Cellular & Developmental Biology. Animal·Jennifer DuongRobert M Hoffman
Feb 28, 2013·Journal of Cellular Biochemistry·Sumiyuki MiiRobert M Hoffman

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Citations

Jul 28, 2017·Cell Cycle·Yasuyuki Amoh, Robert M Hoffman

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Methods Mentioned

BETA
confocal microscopy
transgenic
PCR
electrophoresis

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