Isolation, culture and growth characteristics of dermal papilla cells from Rex rabbits

Liu, Gongyan; Bai, Liya; Li, Shu; Liu, Hongli; Zhu, Yanli; Sun, Haitao; Gao, Shuxia; Jiang, Wenxue; Li, Fuchang

doi:10.1016/j.tice.2020.101348

“…Selected animals were electrically stunned (120 V, pulsed direct current, 50 Hz for 5 s) and euthanized by exsanguination of the carotid artery and before skin was harvested from the experimental rabbits. The DPCs were separated and cultured, methods according to references [51].…”

Section: Animal and Sample Collectionmentioning

confidence: 99%

“…The adenovirus-transfected skin was observed under a positive fluorescence microscope (Nikon ECLIPSE 80i, Japan).Assessment of the proliferation, cell cycle and apoptosis of DPCsDPCs were plated in a 96-well plate at a density of 10 4 cells/well, cultured in basal medium for 24 h, and then transfected with the indicated adenoviruses. The methods assessment of the proliferation according to references[51]. Third-generation DPCs were plated in a disposable 6-well plate at a density of 10 4 cells/mL, with 2 mL of the cell suspension plated in each well.…”

mentioning

confidence: 99%

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

Liu

¹

,

Li

²

,

Liu

³

et al. 2020

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Background: Hair follicles is an appendage from the vertebrate skin epithelium, and arise from the embryonic ectoderm andregenerate cyclically during adult life. Dermal papilla cells (DPCs) is the key dermal component of the hair follicle that directly regulates hair follicle development, growth and regeneration. Recent studies have reported that miRNA plays an important role in regulating hair follicle morphogenesis, proliferation, differentiation and apoptosis of hair follicle stem cells. Results: The miRNAs expression profile of the DPCs from different hair density Rex rabbits shown that 240 differentially expressed of miRNAs were screened (log 2 (HD/LD)｜>1.00 and Q-value≤0.001). Among them, the expression of ocu-miR-205-5p in low hair densities DPCs was higher than that in high hair densities, and it is highly expressed in the skin tissue of Rex rabbits ( P <0.05). ocu-miR-205 could increase cell proliferation and cell apoptosis ratio, change cell cycle process ( P <0.05), affect the genes expression of PI3K/Akt, Wnt, Notch and BMP signaling pathways in DPCs and skin tissue of Rex rabbits, inhibit the protein phosphorylation level of CTNNB1, GSK-3β and the protein expression level of noggin (NOG), promote Akt phosphorylation level ( P <0.05). There was no significant change in primary follicle density ( P >0.05), but the secondary follicle density and total follicle density ( P <0.05) were changed after ocu-miR-205-5p interfered expression, and secondary/primary ratio (S/P) in ocu-miR-205-5p interfered expression group increased at 14 days after injection ( P <0.05). Conclusion: ocu-miR-205 could promote the apoptosis of DPCs, affect PI3K/Akt, Wnt, Notch and BMP signaling pathways genes and proteins expression in DPCs and skin of Rex rabbits, promote the transformation of hair follicles from growth phase to regression and resting phase, and affect hair density of Rex rabbits.

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“…DPCs were plated in a 96-well plate at a density of 10 4 cells/well, cultured in basal medium for 24 h, and then transfected with the indicated adenoviruses. The methods assessment of the proliferation according to references [51]. Third-generation DPCs were plated in a disposable 6-well plate at a density of 10 4 cells/mL, with 2 mL of the cell suspension plated in each well.…”

Section: Assessment Of the Proliferation Cell Cycle And Apoptosis Ofmentioning

confidence: 99%

“…The methods assessment of the cell cycle according to references [51]. For apoptosis assessment, cells were centrifuged and collected into a 1.5 mL centrifugal tube, and then washed with PBS.…”

Section: Assessment Of the Proliferation Cell Cycle And Apoptosis Ofmentioning

confidence: 99%

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

Liu

¹

,

Li

²

,

Liu

³

et al. 2020

Preprint

Self Cite

0

View full text Add to dashboard Cite

Background: Hair follicles is an appendage from the vertebrate skin epithelium, and arise from the embryonic ectoderm andregenerate cyclically during adult life. Dermal papilla cells (DPCs) is the key dermal component of the hair follicle that directly regulates hair follicle development, growth and regeneration. Recent studies have reported that miRNA plays an important role in regulating hair follicle morphogenesis, proliferation, differentiation and apoptosis of hair follicle stem cells. Results: The miRNAs expression profile of the DPCs from different hair density Rex rabbits shown that 240 differentially expressed of miRNAs were screened (log 2 (HD/LD)｜>1.00 and Q-value≤0.001). Among them, the expression of ocu-miR-205-5p in low hair densities DPCs was higher than that in high hair densities, and it is highly expressed in the skin tissue of Rex rabbits ( P <0.05). ocu-miR-205 could increase cell proliferation and cell apoptosis ratio, change cell cycle process ( P <0.05), affect the genes expression of PI3K/Akt, Wnt, Notch and BMP signaling pathways in DPCs and skin tissue of Rex rabbits, inhibit the protein phosphorylation level of CTNNB1, GSK-3β and the protein expression level of noggin (NOG), promote Akt phosphorylation level ( P <0.05). There was no significant change in primary follicle density ( P >0.05), but the secondary follicle density and total follicle density ( P <0.05) were changed after ocu-miR-205-5p interfered expression, and secondary/primary ratio (S/P) in ocu-miR-205-5p interfered expression group increased at 14 days after injection ( P <0.05). Conclusion: ocu-miR-205 could promote the apoptosis of DPCs, affect PI3K/Akt, Wnt, Notch and BMP signaling pathways genes and proteins expression in DPCs and skin of Rex rabbits, promote the transformation of hair follicles from growth phase to regression and resting phase, and affect hair density of Rex rabbits.

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“…The tissues were then minced with ophthalmological scissors and digested with 0.1 mg/ml Collagenase D (Sigma, Cat. C2674, USA) for 6 h at 37 °C, when the dissociation of DPCs from liquid fat was observed under the microscope (Nikon Ts2FL, Japan)[51,52].Subsequently, DPCs were purified by a series of washes with PBS till the supernatant was completely clear, and it was then filtered with a 75 mM filter to remove single cells. Following purification, DPCs were trypsinized to obtain single cells that were then cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, USA) containing 10 % foetal bovine serum (FBS; Gibco, USA) at 37 °C in a humidified atmosphere composed of 95 % air and 5 % CO2.…”

mentioning

confidence: 99%

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

Liu

¹

,

Li

²

,

Liu

³

et al. 2020

Preprint

Self Cite

0

View full text Add to dashboard Cite

Background: Hair follicles is an appendage from the vertebrate skin epithelium, and arise from the embryonic ectoderm andregenerate cyclically during adult life. Dermal papilla cells (DPCs) is the key dermal component of the hair follicle that directly regulates hair follicle development, growth and regeneration. Recent studies have reported that miRNA plays an important role in regulating hair follicle morphogenesis, proliferation, differentiation and apoptosis of hair follicle stem cells. Results: The miRNAs expression profile of the DPCs from different hair density Rex rabbits shown that 240 differentially expressed of miRNAs were screened (log 2 (HD/LD)｜>1.00 and Q-value≤0.001). Among them, the expression of ocu-miR-205-5p in low hair densities DPCs was higher than that in high hair densities, and it is highly expressed in the skin tissue of Rex rabbits ( P <0.05). ocu-miR-205 could increase cell proliferation and cell apoptosis ratio, change cell cycle process ( P <0.05), affect the genes expression of PI3K/Akt, Wnt, Notch and BMP signaling pathways in DPCs and skin tissue of Rex rabbits, inhibit the protein phosphorylation level of CTNNB1, GSK-3β and the protein expression level of noggin (NOG), promote Akt phosphorylation level ( P <0.05). There was no significant change in primary follicle density ( P >0.05), but the secondary follicle density and total follicle density ( P <0.05) were changed after ocu-miR-205-5p interfered expression, and secondary/primary ratio (S/P) in ocu-miR-205-5p interfered expression group increased at 14 days after injection ( P <0.05). Conclusion: ocu-miR-205 could promote the apoptosis of DPCs, affect PI3K/Akt, Wnt, Notch and BMP signaling pathways genes and proteins expression in DPCs and skin of Rex rabbits, promote the transformation of hair follicles from growth phase to regression and resting phase, and affect hair density of Rex rabbits.

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Isolation, culture and growth characteristics of dermal papilla cells from Rex rabbits

Cited by 11 publications

References 34 publications

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits

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