Comparison of two cellular harvesting methods for primary human oral culture of keratinocytes

Klingbeil, María Fátima Guarizo; Herson, Marisa Roma; Cristo, Elier B.; Pinto, Décio dos Santos; Yoshito, Daniele; Mathor, Mônica Beatriz

doi:10.1007/s10561-009-9122-7

Cited by 23 publications

(21 citation statements)

References 17 publications

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“…The average initial harvesting time for keratinocyte cultivation by the direct explant technique was found to be14.2 days in a previous study (7). In the current study, it was 20.25±1.05 days, somewhat longer than the previous study.…”

Section: Discussioncontrasting

confidence: 45%

See 1 more Smart Citation

Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

Bayar¹,

Aydıntuğ²,

Günhan³

et al. 2012

Balkan Med J

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Objective: The aim of this study is the histological and immunohistochemical evaluation of ex vivo produced oral mucosal equivalents using keratinocytes cultured by direct explant technique.Material and Methods: Oral mucosa tissue samples were obtained from the keratinized gingival tissues of 14 healthy human subjects. Human oral mucosa keratinocytes from an oral mucosa biopsy specimen were dissociated by the explant technique. Once a sufficient population of keratinocytes was reached, they were seeded onto the type IV collagen coated "AlloDerm" and taken for histological and immunohistochemical examinations at 11 days postseeding of the keratinocytes on the cadaveric human dermal matrix.Results: Histopathologically and immunohistochemically, 12 out of 14 successful ex vivo produced oral mucosa equivalents (EVPOME) that consisted of a stratified epidermis on a dermal matrix have been developed with keratinocytes cultured by the explant technique. Conclusion:The technical handling involved in the direct explant method at the beginning of the process has fewer steps than the enzymatic method and use of the direct explant technique protocol for culturing of human oral mucosa keratinocyte may be more adequate for EVPOME production.

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Section: Discussioncontrasting

confidence: 45%

“…Basically, there are two techniques used in cultivation of oral mucosa keratinocytes: the enzymatic and the direct explant technique (6,7).…”

Section: Introductionmentioning

confidence: 99%

Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

Bayar¹,

Aydıntuğ²,

Günhan³

et al. 2012

Balkan Med J

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“…Primary explant culture is another approach to isolate cells from various tissues (9–11). A simple small fragment of tissue that adheres to the growth surface will usually give rise to an outgrowth of cells.…”

mentioning

confidence: 99%

Explant Culture: An Efficient Method to Isolate Adipose‐Derived Stromal Cells for Tissue Engineering

et al. 2010

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Enzymatic digestion, the commonly used method of adipose-derived stromal cells isolation, is time consuming and expensive, especially when applied to large volumes of tissue. In the present study, the characteristics of the cells obtained by adipose tissue explant culture were studied. We found that adipose tissue fragments could adhere onto the growth surface of flasks in a very short time after plating and that fibroblast-like cells migrated from the explants and reached confluence. Morphologic analysis and surface markers expression suggested the mesenchymal origin of the cells derived from adipose tissue explants. After in vitro expansion these cells were successfully induced into adipogenic, osteogenic, and chondrogenic lineages, which demonstrated their multipotency. The high growth rate and colony-forming efficiency of explant-derived cells were similar to those of cells obtained by digestion. Furthermore, explant culture gave higher yield of cells than digestion method after primary culture. The experiment of ectopic adipogenesis in nude mice suggested the prospects for tissue engineering of these cells. In conclusion, we obtained multipotent stromal cells from adipose tissue by explant culture, and this method was simple, time saving, and gave a high yield of cells. Therefore, explant culture can be used as an effective way to isolate adipose-derived stromal cells for tissue engineering.

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“…OSCC as well as HaCat cell lines were grown as described previously (25). Normal oral keratinocytes (NOK) were obtained from oral epithelial fragments under enzymatic digestion method, kindly provided by Dr Maria Fatima Guarizo Klingbeil (29). …”

Section: Methodsmentioning

confidence: 99%

Homeobox gene expression profile indicates HOXA5 as a candidate prognostic marker in oral squamous cell carcinoma

Rodini¹,

Xavier²,

Paiva³

et al. 2011

International Journal of Oncology

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The search for molecular markers to improve diagnosis, individualize treatment and predict behavior of tumors has been the focus of several studies. This study aimed to analyze homeobox gene expression profile in oral squamous cell carcinoma (OSCC) as well as to investigate whether some of these genes are relevant molecular markers of prognosis and/or tumor aggressiveness. Homeobox gene expression levels were assessed by microarrays and qRT-PCR in OSCC tissues and adjacent non-cancerous matched tissues (margin), as well as in OSCC cell lines. Analysis of microarray data revealed the expression of 147 homeobox genes, including one set of six at least 2-fold up-regulated, and another set of 34 at least 2-fold down-regulated homeobox genes in OSCC. After qRT-PCR assays, the three most up-regulated homeobox genes (HOXA5, HOXD10 and HOXD11) revealed higher and statistically significant expression levels in OSCC samples when compared to margins. Patients presenting lower expression of HOXA5 had poorer prognosis compared to those with higher expression (P=0.03). Additionally, the status of HOXA5, HOXD10 and HOXD11 expression levels in OSCC cell lines also showed a significant up-regulation when compared to normal oral keratinocytes. Results confirm the presence of three significantly upregulated (>4-fold) homeobox genes (HOXA5, HOXD10 and HOXD11) in OSCC that may play a significant role in the pathogenesis of these tumors. Moreover, since lower levels of HOXA5 predict poor prognosis, this gene may be a novel candidate for development of therapeutic strategies in OSCC.

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Comparison of two cellular harvesting methods for primary human oral culture of keratinocytes

Cited by 23 publications

References 17 publications

Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

Explant Culture: An Efficient Method to Isolate Adipose‐Derived Stromal Cells for Tissue Engineering

Homeobox gene expression profile indicates HOXA5 as a candidate prognostic marker in oral squamous cell carcinoma

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