Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cationic currents in mature MDCK cells are almost exclusively due to K+ channels. Harvesting with trypsin-EDTA destroys 80-90% of these channels. Upon replating, K+ currents recover in 12-20 h, by means a process that requires synthesis of proteins and of RNA. In the present work we demonstrate that this restoration depends on a Ca2+ activated-cell contact. Thus, cells in confluent monolayers bathed with 1.8 mM Ca2+ have a K+ current of 343 +/- 82 pA; confluent without Ca2+ have only 90 +/- 12 pA (27% of control; and without cell-cell contacts incubated with 1.8 mM Ca2+ (subconfl+Ca2+) have 104 +/- 21 pA (31% of control). This demonstration that the expression of K+ channels depends on Ca-activated cell-attaching molecules suggests that a molecule of the type of uvomorulin is involved.
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PMID:[The role of cellular contacts in the recovery of the ion channels of epithelial cell membranes]. 925 81

Human skin has continuous self-renewal potential throughout adult life and serves as first line of defence. Its cellular components such as human epidermal keratinocytes (HEKs) and dermal mesenchymal stromal cells (DMSCs) are valuable resources for wound healing applications and cell based therapies. Here we show a simple, scalable and cost-effective method for sequential isolation and propagation of HEKs and DMSCs under defined culture conditions. Human skin biopsy samples obtained surgically were cut into fine pieces and cultured employing explant technique. Plated skin samples attached and showed outgrowth of HEKs. Gross microscopic examination displayed polygonal cells with a granular cytoplasm and H&E staining revealed archetypal HEK morphology. RT-PCR and immunocytochemistry authenticated the presence of key HEK markers including trans-membrane protein epithelial cadherin (E-cadherin), keratins and cytokeratin. After collection of HEKs by trypsin-EDTA treatment, mother explants were left intact and cultured further. Interestingly, we observed the appearance of another cell type with fibroblastic or stromal morphology which were able to grow up to 15 passages in vitro. Growth pattern, expression of cytoskeletal protein vimentin, surface proteins such as CD44, CD73, CD90, CD166 and mesodermal differentiation potential into osteocytes, adipocytes and chondrocytes confirmed their bonafide mesenchymal stem cell like status. These findings albeit preliminary may open up significant opportunities for novel applications in wound healing.
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PMID:Sequential cultivation of human epidermal keratinocytes and dermal mesenchymal like stromal cells in vitro. 2569 60


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