Limits...
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.

Show MeSH

Related in: MedlinePlus

DAPI nuclear staining and P75NTR immuostaining of cryopreserved or fresh follicles after 26 days in Gelfoam histoculture.(A) DAPI nuclear staining. Fresh hair follicles maintained intact structures of the hair shaft, inner root sheath, and outer root sheath, with a large amount of nestin-GFP HAP stem cells growing under the bulb area after 26 days of Gelfoam histoculture. In contrast, the inner structures of Gelfoam-histocultured hair follicles after DMSO- or glycerol-cryopreservation had no DAPI-stained nuclei, indicating damage of the inner structure of the follicle after cryopreservation. Most of the nestin-GFP expressing HAP stem cells grew around the outer root sheath and bulb area in cryopreserved hair follicles. (B) p75NTR immunostaining of frozen sections colocalized with ND-GFP HAP stem cells. These results indicate that ND-GFP HAP stem cells in cryopreserved follicles were maintained.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4696652&req=5

pone.0145997.g003: DAPI nuclear staining and P75NTR immuostaining of cryopreserved or fresh follicles after 26 days in Gelfoam histoculture.(A) DAPI nuclear staining. Fresh hair follicles maintained intact structures of the hair shaft, inner root sheath, and outer root sheath, with a large amount of nestin-GFP HAP stem cells growing under the bulb area after 26 days of Gelfoam histoculture. In contrast, the inner structures of Gelfoam-histocultured hair follicles after DMSO- or glycerol-cryopreservation had no DAPI-stained nuclei, indicating damage of the inner structure of the follicle after cryopreservation. Most of the nestin-GFP expressing HAP stem cells grew around the outer root sheath and bulb area in cryopreserved hair follicles. (B) p75NTR immunostaining of frozen sections colocalized with ND-GFP HAP stem cells. These results indicate that ND-GFP HAP stem cells in cryopreserved follicles were maintained.

Mentions: After 26 days in Gelfoam histoculture, 3D confocal images showed that ND-GFP HAP stem cell outgrowth occurred both at the upper and lower part of fresh hair follicles (Fig 2C). DAPI nuclear staining showed that the fresh hair follicles maintained normal structures in the hair shaft, inner root sheath, and outer root sheath, and a large amount of ND-GFP HAP stem cells grew around the bulb area (Fig 3A). In DMSO cryopreserved follicles, ND-GFP 3D images indicated similar HAP stem cells outgrowth both in the upper and lower part of hair follicle, but with less cell outgrowth than in fresh hair follicles (Fig 2C). With glycerol-cryopreservation, the recovered hair follicles had less ND-GFP HAP stem cells. The cultured hair follicles had ND-GFP HAP stem cells mostly in the outer root sheath and the bulb area after both DMSO- and glycerol-cryopreservation. In contrast, the inner structure of cultured hair follicles had little DAPI nuclear staining remained after either method of cryopreservation. These results indicate cell death of the inner part and structural damage of the frozen hair follicle in both cryopreservation media (Fig 3A). The bulb area of the hair follicle had ND-GFP-nestin HAP stem cells growing out, and the dermal papilla area maintained DAPI nuclear staining after both cryopreservation methods. These results suggest the survival of bulb area and dermal papilla cells in both DMSO and glycerol-cryopreserved hair follicles (Fig 3B). p75NTR immunostaining also indicated ND-GFP HAP stem cells were maintained in both conditions of cryopreservation (Fig 3B).


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)

DAPI nuclear staining and P75NTR immuostaining of cryopreserved or fresh follicles after 26 days in Gelfoam histoculture.(A) DAPI nuclear staining. Fresh hair follicles maintained intact structures of the hair shaft, inner root sheath, and outer root sheath, with a large amount of nestin-GFP HAP stem cells growing under the bulb area after 26 days of Gelfoam histoculture. In contrast, the inner structures of Gelfoam-histocultured hair follicles after DMSO- or glycerol-cryopreservation had no DAPI-stained nuclei, indicating damage of the inner structure of the follicle after cryopreservation. Most of the nestin-GFP expressing HAP stem cells grew around the outer root sheath and bulb area in cryopreserved hair follicles. (B) p75NTR immunostaining of frozen sections colocalized with ND-GFP HAP stem cells. These results indicate that ND-GFP HAP stem cells in cryopreserved follicles were maintained.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0145997.g003: DAPI nuclear staining and P75NTR immuostaining of cryopreserved or fresh follicles after 26 days in Gelfoam histoculture.(A) DAPI nuclear staining. Fresh hair follicles maintained intact structures of the hair shaft, inner root sheath, and outer root sheath, with a large amount of nestin-GFP HAP stem cells growing under the bulb area after 26 days of Gelfoam histoculture. In contrast, the inner structures of Gelfoam-histocultured hair follicles after DMSO- or glycerol-cryopreservation had no DAPI-stained nuclei, indicating damage of the inner structure of the follicle after cryopreservation. Most of the nestin-GFP expressing HAP stem cells grew around the outer root sheath and bulb area in cryopreserved hair follicles. (B) p75NTR immunostaining of frozen sections colocalized with ND-GFP HAP stem cells. These results indicate that ND-GFP HAP stem cells in cryopreserved follicles were maintained.
Mentions: After 26 days in Gelfoam histoculture, 3D confocal images showed that ND-GFP HAP stem cell outgrowth occurred both at the upper and lower part of fresh hair follicles (Fig 2C). DAPI nuclear staining showed that the fresh hair follicles maintained normal structures in the hair shaft, inner root sheath, and outer root sheath, and a large amount of ND-GFP HAP stem cells grew around the bulb area (Fig 3A). In DMSO cryopreserved follicles, ND-GFP 3D images indicated similar HAP stem cells outgrowth both in the upper and lower part of hair follicle, but with less cell outgrowth than in fresh hair follicles (Fig 2C). With glycerol-cryopreservation, the recovered hair follicles had less ND-GFP HAP stem cells. The cultured hair follicles had ND-GFP HAP stem cells mostly in the outer root sheath and the bulb area after both DMSO- and glycerol-cryopreservation. In contrast, the inner structure of cultured hair follicles had little DAPI nuclear staining remained after either method of cryopreservation. These results indicate cell death of the inner part and structural damage of the frozen hair follicle in both cryopreservation media (Fig 3A). The bulb area of the hair follicle had ND-GFP-nestin HAP stem cells growing out, and the dermal papilla area maintained DAPI nuclear staining after both cryopreservation methods. These results suggest the survival of bulb area and dermal papilla cells in both DMSO and glycerol-cryopreserved hair follicles (Fig 3B). p75NTR immunostaining also indicated ND-GFP HAP stem cells were maintained in both conditions of cryopreservation (Fig 3B).

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