Human dermal reticular fibroblasts at confluence display a signature micro pattern <i>in vitro</i>

Abstract

This paper sets out to demonstrate that scraping of the flat dorsal surface of human dermis with a scalpel blade and cell plating without centrifugation can lead to the recognition and identification of the individual packing micro pattern of dermal reticular fibroblasts at confluence. The characteristic alignment of papillary and reticular fibroblasts at right angles to each other led to the positive identification of reticular fibroblasts. A non-enzymatic means of sub-culturing (passaging), which yields fully functional, healthy cells with normal, phenotypic morphology is also described. Implications for published subcutaneous wound healing studies are discussed as well as the confluent reticular fibroblast configuration, interpreted as an anatomic site identity code, which may be the address of a specific fibroblast gene pattern expression.

Scientific Viewpoints with Emphasis on Dermal Cellular Regeneration in Wound Sites

“…Firstly, pure reticular fibroblast cell isolation has been a scientific problem. Unless a segment of skin tissue (after stripping off the epidermis with a pair of forceps) was specifically cut with a dermatome, and followed by enzyme use or the setting up of a classic "explant" cell culture, scientists would obtain a mixture of papillary and reticular fibroblasts after enzyme use on minced dermis or the loose spread of fibroblasts from the explant which lacked the side-on packing configuration at an angled vertical earlier described for primary reticular fibroblasts at confluence [11]. For years, clinicians have treated the wound site with a variety of dressings which have become more sophisticated and expensive e.g.…”

“…The procedure below can be used separately with a monolayer of papillary or reticular fibroblasts at confluence [11], but instead of 20% FBS, 10% autologous plasma will be used with (or without) the culture medium (IMDM, Iscove's modified Dulbecco's medium or Medium 199). A cell scraper or if sub-culturing, 5 mM EDTA/DPBS pH 7.4 on confluent monolayers may be used.…”

“…Time to confluence will be monitored. However, for the time being, a straight forward swop of 10% plasma for the normal 20% foetal bovine serum (FBS) [11] is being put forward for use. The percentage of plasma will be used with or without the culture medium to determine the optimal criteria.…”

“…Pressing a finger into a former wound site reveals a depression which can be not immediately apparent at first sight. This author has recently described the signature side-on packing configuration at confluence of human dermal reticular fibroblasts at an angled vertical in vitro, obtained by non-enzymatic isolation, using 20% foetal bovine serum (FBS) [11]. These cell cultures can be established by using a patient's own plasma.…”

Tackling Post-Operative Cutaneous Scarring with Autologous Cell Therapy

“…This author has demonstrated, albeit in an in vitro setting, that un-manipulated primary (P 0 ) human dermal reticular fibroblasts retain their spatial configuration and their signature packing arrangement at an angle to the vertical [7]. Whether these cells have a short-or long-term memory has not been determined.…”