One-step collagenase I treatment: an efficient way for isolation and cultivation of human scalp dermal papilla cells

Yu, Li; Li, Gou Qiang; Lin, Chang Min; Cai, Xianbin

doi:10.1016/j.jdermsci.2004.10.001

Cited by 22 publications

(19 citation statements)

References 6 publications

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“…DPCs were isolated from human scalp tissue and subjected to primary culture and subculture [12]. Human scalp tissues were obtained with informed consent from the First Affiliated Hospital, Shantou University Medical College.The protocols used in the study were approved by the Hospital’s Biomedical Ethics Committee (permit number 2015–42), and all participants provided written informed consent.…”

Section: Methodsmentioning

confidence: 99%

iTRAQ-Based Quantitative Proteomic Comparison of Early- and Late-Passage Human Dermal Papilla Cell Secretome in Relation to Inducing Hair Follicle Regeneration

et al. 2016

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Alopecia is an exceedingly prevalent problem that lacks effective therapy. Recently, research has focused on early-passage dermal papilla cells (DPCs), which have hair inducing activity both in vivo and in vitro. Our previous study indicated that factors secreted from early-passage DPCs contribute to hair follicle (HF) regeneration. To identify which factors are responsible for HF regeneration and why late-passage DPCs lose this potential, we collected 48-h-culture medium (CM) from both of passage 3 and 9 DPCs, and subcutaneously injected the DPC-CM into NU/NU mice. Passage 3 DPC-CM induced HF regeneration, based on the emergence of a white hair coat, but passage 9 DPC-CM did not. In order to identify the key factors responsible for hair induction, CM from passage 3 and 9 DPCs was analyzed by iTRAQ-based quantitative proteomic technology. We identified 1360 proteins, of which 213 proteins were differentially expressed between CM from early-passage vs. late-passage DPCs, including SDF1, MMP3, biglycan and LTBP1. Further analysis indicated that the differentially-expressed proteins regulated the Wnt, TGF-β and BMP signaling pathways, which directly and indirectly participate in HF morphogenesis and regeneration. Subsequently, we selected 19 proteins for further verification by multiple reaction monitoring (MRM) between the two types of CM. These results indicate DPC-secreted proteins play important roles in HF regeneration, with SDF1, MMP3, biglycan, and LTBP1 being potential key inductive factors secreted by dermal papilla cells in the regeneration of hair follicles.

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Section: Methodsmentioning

confidence: 99%

iTRAQ-Based Quantitative Proteomic Comparison of Early- and Late-Passage Human Dermal Papilla Cell Secretome in Relation to Inducing Hair Follicle Regeneration

et al. 2016

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“…Collagenase has been used previously for enhancing isolation of HF stem cells for cultured expansion. 17 The ORS, dermal sheath and papilla are a reservoir for melanocytes and stem cells (including mesenchymal, epithelial and melanocyte stem cells). Often, the latter are poorly defined, heterogeneous populations that lack a single specific identifying marker.…”

Section: Discussionmentioning

confidence: 99%

A randomized pilot study to compare hair follicle cell suspensions prepared using trypsin alone versus trypsin in combination with collagenase type I for transplantation in vitiligo

et al. 2019

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Summary Background Noncultured extracted hair follicle outer root sheath cell suspension (NC‐EHF‐ORS‐CS) is an upcoming surgical technique to treat stable vitiligo. Conventionally it employs trypsin to tap the hair follicle (HF) reservoir for autologous melanocytes and their precursors for transplantation. However, a perifollicular dermal sheath composed of type 1 collagen encases the target ‘bulge’ region of the HF. Adding collagenase type 1 would digest the ORS, facilitating better release of cells. Aim To compare the repigmentation achieved using trypsin and a combination of collagenase plus trypsin, respectively, with dermabrasion alone, and to compare cell counts, viability and composition of both suspensions. Methods This was a randomized, double‐blind, comparative, therapeutic trial, conducted as a pilot study on 22 patients with stable vitiligo. Three similar patches were randomized into three parallel treatment arms [(A) trypsin plus collagenase, (B) trypsin alone and (C) dermabrasion with vehicle alone]. Each patient’s HF sample was divided and digested by the two methods, and transplanted as suspensions onto dermabraded patches, while a third dermabraded patch received the vehicle only. Suspensions were sent for laboratory analysis. Repigmentation was assessed over a follow‐up of 6 months. Results There was a significant increase in cell yield and comparable viability when collagenase was added. Immunohistochemical and flow cytometry studies showed a nonsignificant increase in HMB45+ melanocytes and their precursor stem cells in group A. This trend was reflected clinically in the extent of repigmentation [group A (33.22%) > B (24.31%) > C (16.59%); P = 0.13]. Adding collagenase induced significantly higher repigmentation than dermabrasion alone (P < 0.05). Conclusions Incorporating collagenase type I into the conventional NC‐EHF‐ORS‐CS technique resulted in enhanced retrieval of pigment‐forming cells and subsequently improved repigmentation in vitiligo.

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“…Isolation of DPCs was performed according to the method described by Li et al . Briefly, DPCs were digested in type IV collagenase (Sigma) from the bulbs of hair follicles, randomly transferred onto 24‐well plates (BD Biosciences) which were coated with Matrigel, hyaluronan, type IV collagen or fibronectin, and cultured in Dulbecco modified Eagle medium (DMEM) (Gibco, Grand Island, NY, USA) supplemented with 20% (v/v) fetal bovine serum (FBS) (Gibco) at 37 °C in a humidified 5% CO 2 atmosphere.…”

Section: Methodsmentioning

confidence: 99%

Effects of extracellular matrix on the growth characteristics of human dermal papillae cellsin vitro

Miao

Fan

et al. 2016

Clin Exp Dermatol

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One-step collagenase I treatment: an efficient way for isolation and cultivation of human scalp dermal papilla cells

Cited by 22 publications

References 6 publications

iTRAQ-Based Quantitative Proteomic Comparison of Early- and Late-Passage Human Dermal Papilla Cell Secretome in Relation to Inducing Hair Follicle Regeneration

iTRAQ-Based Quantitative Proteomic Comparison of Early- and Late-Passage Human Dermal Papilla Cell Secretome in Relation to Inducing Hair Follicle Regeneration

A randomized pilot study to compare hair follicle cell suspensions prepared using trypsin alone versus trypsin in combination with collagenase type I for transplantation in vitiligo

Effects of extracellular matrix on the growth characteristics of human dermal papillae cellsin vitro

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