Mucosal keratinocyte isolation: a short comparative study on thermolysin and dispase

Rakhorst, Hinne; Tra, W.; Sluijs, Sandra J. Posthumus-van; Groot, Eveline; Osch, Gerjo J.V.M. van; Neck, Johan W. van; Hofer, Stefan O. P.

doi:10.1016/j.ijom.2006.06.011

Cited by 10 publications

(8 citation statements)

References 21 publications

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“…However, this method results in the lowest growth rate of isolated keratinocyte (Figure A) that is in line with observations described elsewhere . Furthermore, the effectiveness of thermolysin digestion may be unpredictable and may lead to loss of basal, highly proliferative keratinocytes including stem cells …”

Section: Resultssupporting

confidence: 89%

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Isolation and culture of human primary keratinocytes—a methods review

Ścieżyńska

Nogowska

Sikorska

et al. 2019

Experimental Dermatology

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Keratinocyte culture is a necessary and widely used tool in a variety of experimental dermatological and biomedical studies. Literature search has revealed a variety of different protocols of human primary keratinocyte isolation and culture. Therefore, the aim of this paper was to review and summarize current trends in human primary keratinocyte culture techniques. We present data on the most popular and effective methods of human keratinocyte isolation and cultivation obtained from screening of 945 papers published during the last 10 years.

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Section: Resultssupporting

confidence: 89%

“…Two‐step digestion method appears to decrease the risk of fibroblast contamination and minimize usage of trypsin which in high concentration may affect viability of the keratinocytes …”

Section: Introductionmentioning

confidence: 99%

Isolation and culture of human primary keratinocytes—a methods review

Ścieżyńska

Nogowska

Sikorska

et al. 2019

Experimental Dermatology

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“…The culture of RCECs was performed as described previously [31]. Briefly, RCECs were cultured in Keratinocyte-SFM (1∶1) medium (Gibco) containing 5% fetal bovine serum (FBS), 0.05 mg/ml bovine pituitary extract, 5 ng/ml epidermal growth factor, 100 U/ml penicillin and 100 µg/ml streptomycin.…”

Section: Methodsmentioning

confidence: 99%

Effect of Reactive Oxygen Species Generation in Rabbit Corneal Epithelial Cells on Inflammatory and Apoptotic Signaling Pathways in the Presence of High Osmotic Pressure

Chen

et al. 2013

PLoS ONE

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It is generally accepted that high osmotic pressure (HOP) of lacrimal fluid is the core mechanism causing ocular inflammation and injury. However, the association between HOP and the regulation of cell inflammatory response and apoptotic pathways remains unclear. In the present study, we used HOP to interfere with in vitro cultured rabbit corneal epithelial cells, and found that HOP increased the generation of reactive oxygen species (ROS) in rabbit corneal epithelial cells, and increased ROS in turn induced the activation of JNK inflammatory signaling pathway, which further promoted the expression of pro-inflammatory factor NF-κβ and induced the generation of inflammatory factor IL-1β and TNF-α. In addition, HOP-induced ROS in rabbit corneal epithelial cells regulated the CD95/CD95L-mediated cell apoptotic signaling pathway by activating JNK inflammatory signaling pathway. These findings may serve as new theoretical basis and a new way of thinking about the treatment of ocular diseases, especially dry eye.

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“…These keratinocytes have less proliferative potential than the CK13-negative cells at the basement membrane of the epidermis 6) . The immunohistochemical detection of CK13, which can be observed regularly in non-cornifying squamous epithelia, provides no information for the timeestimation of wounds in human skin, since no significant…”

Section: Introductionmentioning

confidence: 94%

Regeneration of Keratinocytes, Langerhans Cells, Merkel Cells and Melanocytes after Removal of the Palatal Mucosa in the Early Healing Period

明展¹,

賢一²,

克寿³

et al. 2010

J. Jpn. Oral Muco. Membr.

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Abstract：This study investigated the process of wound healing in the palatal mucosa, focusing on the proliferation and differentiation of keratinocytes, and regeneration of the basement membrane, Langerhans cells, Merkel cells and melanocytes. Golden hamsters were used as experimental animals and the entire fourth palatine ruga including the periosteum was removed mechanically with a scalpel. Control and experimental animals were sacrificed 3, 7, 14 and 21 days later. Maxillae were removed and fixed with 10％ formalin. Paraffin sections were then cut and immunohistochemically stained. For immunohistochemical staining, antibodies against PCNA for detecting the proliferating cells, CK14 and CK13 for epithelial differentiation, type IV collagen for the basement membrane, S-100for Langerhans cells and melanocytes, and CK20 for Merkel cells were used. At 14 days, the wound regions were covered with epithelium. CK14 and CK13 staining showed that epithelial differentiation had not been completed either in peripheral or central regions by 21 days. Type IV collagen was detected only in some parts of the epithelial basement membrane on day 14 and 21 in the central region. No S-100 or CK20-positive cell was found in the regenerated epithelium through 21 days in the peripheral regions and the central region. These results suggest that wound healing of the palatal mucosa seems to be macroscopically normal, but normalization of epithelial differentiation and regeneration of the basement membrane require a substantial amount of time. It is also suggested that the immune functions of Langerhans cells and the protective functions of melanocytes cannot regenerate, and that the absence of Merkel cells reduces sensory perception for at least 21 days.

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Mucosal keratinocyte isolation: a short comparative study on thermolysin and dispase

Cited by 10 publications

References 21 publications

Isolation and culture of human primary keratinocytes—a methods review

Isolation and culture of human primary keratinocytes—a methods review

Effect of Reactive Oxygen Species Generation in Rabbit Corneal Epithelial Cells on Inflammatory and Apoptotic Signaling Pathways in the Presence of High Osmotic Pressure

Regeneration of Keratinocytes, Langerhans Cells, Merkel Cells and Melanocytes after Removal of the Palatal Mucosa in the Early Healing Period

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