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Extensive Hair Shaft Growth after Mouse Whisker Follicle Isolation, Cryopreservation and Transplantation in Nude Mice.

Cao W, Li L, Tran B, Kajiura S, Amoh Y, Liu F, Hoffman RM - PLoS ONE (2015)

Bottom Line: DMSO better cryopreserved mouse whisker follicles compared to glycerol.Cryopreserved hair follicles also maintained the hair follicle-associated-pluripotent (HAP) stem cells, evidenced by P75NTR expression.Subcutaneous transplantation of DMSO-cryopreserved hair follicles in nude mice resulted in extensive hair fiber growth over 8 weeks, indicating the functional recovery of hair shaft growth of cryopreserved hair follicles.

View Article: PubMed Central - PubMed

Affiliation: AntiCancer Inc., San Diego, CA, 92111, United States of America.

ABSTRACT
We previously demonstrated that whole hair follicles could be cryopreserved to maintain their stem-cells differentation potential. In the present study, we demonstrated that cryopreserved mouse whisker hair follicles maintain their hair growth potential. DMSO better cryopreserved mouse whisker follicles compared to glycerol. Cryopreserved hair follicles also maintained the hair follicle-associated-pluripotent (HAP) stem cells, evidenced by P75NTR expression. Subcutaneous transplantation of DMSO-cryopreserved hair follicles in nude mice resulted in extensive hair fiber growth over 8 weeks, indicating the functional recovery of hair shaft growth of cryopreserved hair follicles.

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Confocal 3D images of ND-GFP hair-follicle-associated-pluripotent (HAP) stem cells in whisker follicles grown in Gelfoam histoculture.(A) ND-GFP-expressing HAP stem cells in DMSO- or glycerol-cryopreserved hair follicles or fresh follicles at day 2, 10, 16 and 26 days after Gelfoam histoculture. (B) Quantitative analysis of ND-GFP HAP stem cells fluorescence in cultured whisker follicles. DMSO-cryopserved follicles had more extensive growth of HAP stem cells than in glycerol-cryopreserved follicles, but less than in fresh follicles. (C) At day 26 after culture on Gelfoam, only 2/6 hair follicles that were cryopreserved in glycerol recovered. In contrast, 6/6 hair follicles recovered after DMSO cryopreservation, similar to fresh follicles.
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pone.0145997.g002: Confocal 3D images of ND-GFP hair-follicle-associated-pluripotent (HAP) stem cells in whisker follicles grown in Gelfoam histoculture.(A) ND-GFP-expressing HAP stem cells in DMSO- or glycerol-cryopreserved hair follicles or fresh follicles at day 2, 10, 16 and 26 days after Gelfoam histoculture. (B) Quantitative analysis of ND-GFP HAP stem cells fluorescence in cultured whisker follicles. DMSO-cryopserved follicles had more extensive growth of HAP stem cells than in glycerol-cryopreserved follicles, but less than in fresh follicles. (C) At day 26 after culture on Gelfoam, only 2/6 hair follicles that were cryopreserved in glycerol recovered. In contrast, 6/6 hair follicles recovered after DMSO cryopreservation, similar to fresh follicles.

Mentions: After thawing and culture on Gelfoam for 4 weeks, the ND-GFP fluorescence of the HAP stem cells was measured. Confocal real-time 3D images of ND-GFP HAP hair follicle on Gelfoam showed that ND-GFP HAP stem-cell-fluorescence in fresh hair follicle had dramatic increases at 10, 16 and 26 days. The increases in HAP stem cell ND-GFP fluorescence after DMSO cryopreservation of hair follicles was slower than in fresh follicles. Glycerol-cryopreserved follicles had less increases in ND-GFP fluorescence than the DMSO-preserved follicles (Fig 2A and 2B). At day 26, 2/6 hair follicles cryopreserved in glycerol recovered to grow ND-GFP HAP stem cells, 2/6 had poor growth, and 2/6 were consider dead. In contrast, 6/6 hair follicles frozen in DMSO recovered (Fig 2C).


Extensive Hair Shaft Growth after Mouse Whisker Follicle Isolation, Cryopreservation and Transplantation in Nude Mice.

Cao W, Li L, Tran B, Kajiura S, Amoh Y, Liu F, Hoffman RM - PLoS ONE (2015)

Confocal 3D images of ND-GFP hair-follicle-associated-pluripotent (HAP) stem cells in whisker follicles grown in Gelfoam histoculture.(A) ND-GFP-expressing HAP stem cells in DMSO- or glycerol-cryopreserved hair follicles or fresh follicles at day 2, 10, 16 and 26 days after Gelfoam histoculture. (B) Quantitative analysis of ND-GFP HAP stem cells fluorescence in cultured whisker follicles. DMSO-cryopserved follicles had more extensive growth of HAP stem cells than in glycerol-cryopreserved follicles, but less than in fresh follicles. (C) At day 26 after culture on Gelfoam, only 2/6 hair follicles that were cryopreserved in glycerol recovered. In contrast, 6/6 hair follicles recovered after DMSO cryopreservation, similar to fresh follicles.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4696652&req=5

pone.0145997.g002: Confocal 3D images of ND-GFP hair-follicle-associated-pluripotent (HAP) stem cells in whisker follicles grown in Gelfoam histoculture.(A) ND-GFP-expressing HAP stem cells in DMSO- or glycerol-cryopreserved hair follicles or fresh follicles at day 2, 10, 16 and 26 days after Gelfoam histoculture. (B) Quantitative analysis of ND-GFP HAP stem cells fluorescence in cultured whisker follicles. DMSO-cryopserved follicles had more extensive growth of HAP stem cells than in glycerol-cryopreserved follicles, but less than in fresh follicles. (C) At day 26 after culture on Gelfoam, only 2/6 hair follicles that were cryopreserved in glycerol recovered. In contrast, 6/6 hair follicles recovered after DMSO cryopreservation, similar to fresh follicles.
Mentions: After thawing and culture on Gelfoam for 4 weeks, the ND-GFP fluorescence of the HAP stem cells was measured. Confocal real-time 3D images of ND-GFP HAP hair follicle on Gelfoam showed that ND-GFP HAP stem-cell-fluorescence in fresh hair follicle had dramatic increases at 10, 16 and 26 days. The increases in HAP stem cell ND-GFP fluorescence after DMSO cryopreservation of hair follicles was slower than in fresh follicles. Glycerol-cryopreserved follicles had less increases in ND-GFP fluorescence than the DMSO-preserved follicles (Fig 2A and 2B). At day 26, 2/6 hair follicles cryopreserved in glycerol recovered to grow ND-GFP HAP stem cells, 2/6 had poor growth, and 2/6 were consider dead. In contrast, 6/6 hair follicles frozen in DMSO recovered (Fig 2C).

Bottom Line: DMSO better cryopreserved mouse whisker follicles compared to glycerol.Cryopreserved hair follicles also maintained the hair follicle-associated-pluripotent (HAP) stem cells, evidenced by P75NTR expression.Subcutaneous transplantation of DMSO-cryopreserved hair follicles in nude mice resulted in extensive hair fiber growth over 8 weeks, indicating the functional recovery of hair shaft growth of cryopreserved hair follicles.

View Article: PubMed Central - PubMed

Affiliation: AntiCancer Inc., San Diego, CA, 92111, United States of America.

ABSTRACT
We previously demonstrated that whole hair follicles could be cryopreserved to maintain their stem-cells differentation potential. In the present study, we demonstrated that cryopreserved mouse whisker hair follicles maintain their hair growth potential. DMSO better cryopreserved mouse whisker follicles compared to glycerol. Cryopreserved hair follicles also maintained the hair follicle-associated-pluripotent (HAP) stem cells, evidenced by P75NTR expression. Subcutaneous transplantation of DMSO-cryopreserved hair follicles in nude mice resulted in extensive hair fiber growth over 8 weeks, indicating the functional recovery of hair shaft growth of cryopreserved hair follicles.

Show MeSH
Related in: MedlinePlus