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Radiation effects on a 3D cell culture of primary human keratinocytes and the protective effect of N-acetyl-cysteine (NAC) and the Traditional Chinese Medicine Qingre-Liyan decoction (QYD)

ABSTRACT: We studied the radiation effect on cell culture model of oral mucositis. The musositis model (tissues) used is an organotypic model which consisted of 3-dimensional (3-D) cell cultures of primary human oral keratinocytes. The tissues were purchased from MatTek corporation (Ashland, MA) and were grown on top of microporous membrane and supplemented with MatTek serum free media. These tissues were irradiated with 12 Gy. Six hours after the irradiation the tissues were cut in half. Half of the tissue was used to extract total RNA (RNeasy Plus Mini kit, Qiagen, Germantown, MD) and the other half was used to evaluate the histology and the apoptosis. We had the following groups: Non-Irradiated Tissue, Irradiated Tissue with 12 Gy; Irradiated Tissue with 12 Gy pretreated with N-acetylcysteine (NAC); Irradiated Tissue with 12 Gy pretreated with the Traditional Chinese Medicine Qingre-Liyan decoction (QYD); and Irradiated Tissue with 12 Gy pretreated with 50:50 w/w NAC-QYD.

ORGANISM(S): Homo sapiens

PROVIDER: GSE62397 | GEO | 2014/10/16

SECONDARY ACCESSION(S): PRJNA264014

REPOSITORIES: GEO

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Radiation effects on a 3D cell culture of primary human keratinocytes and the protective effect of N-acetyl-cysteine and the Traditional Chinese Medicine (TCM) Qingre-Liyan decoction

Project description:We studied the radiation effect on cell culture model of oral mucositis. The musositis model (tissues) used is an organotypic model which consisted of 3-dimensional (3-D) cell cultures of primary human oral keratinocytes. The tissues were purchased from MatTek corporation (Ashland, MA) and were grown on top of microporous membrane and supplemented with MatTek serum free media. These tissues were irradiated with 12 Gy. Six hours after the irradiation the tissues were cut in half. Half of the tissue was used to extract total RNA (RNeasy Plus Mini kit, Qiagen, Germantown, MD) and the other half was used to evaluate the histology and the apoptosis. We had the following groups: Non-Irradiated Tissue, Irradiated Tissue with 12 Gy; Irradiated Tissue with 12 Gy pretreated with N-acetylcysteine (ACS); Irradiated Tissue with 12 Gy pretreated with the Traditional Chinese Medicine (TCM) Qingre-Liyan decoction; and Irradiated Tissue with 12 Gy pretreated with 50:50 w/w ACS:TCM (AT) Group 1: Human Prirmary 3-D Cultures of Human Keratinocytes were irradiated with 12 Gy; Group 2: Human Prirmary 3-D Cultures of Human Keratinocytes were pretereated with N-acetylcysteine (ACS) and then irradiated with 12 Gy; Human Prirmary 3-D Cultures of Human Keratinocytes were pretreated with the Traditional Chinese Medicine (TCM), Qingre-Liyan Decostion and then irradiated with 12 Gy; Human Prirmary 3-D Cultures of Human Keratinocytes were pretreated with a 50:50 w/w mixture of ACS:TCM (AT) and then irradiated with 12 Gy. Each sample was done in triplicate.

2014-10-16 | E-GEOD-62397 | biostudies-arrayexpress

Radiation Effects on a 3-Dimensional Model of Oral Mucositis

Project description:We studied the radiation effect on cell culture model of oral mucositis. The musositis model (tissues) used is an organotypic model which consisted of 3 dimensional (3-D) cell cultures of primary human oral keratinocytes grown on top of primary oral fibroblasts and resembles oral mucosal (tissues). The tissues were purchased from MatTek corporation (Ashland, MA) and were grown on top of microporous membrane and supplemented with MatTek serum free media. These tissues were irradiated with 12 and 2 Gy. Six hours after the irradiation the tissues were cut in half. Half of the tissue was used to extract total RNA (RNeasy Plus Mini kit, Qiagen, Germantown, MD) and the other half was used to evaluate the histology and the apoptosis. We had the following groups: Non-Irradiated Tissue, Irradiated Tissue with 12 Gy and Irradiated Tissues with 2 Gy.

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Project description:We performed bulk RNA sequencing analysis of a 3D organoleptic model of human oral mucosa treated with everolimus. The 3D model (tissue) consisted of human normal keratinocytes grown on top of human fibroblasts and was histologically similar to buccal mucosa. The tissue was purchased from MatTek Corporation (Ashland, MA) and cultured in serum free media. The 3D model was maintained for different times in the tissue culture (40 hours and 60 hours from the time received) and treated with different concentration of Everolimus (no drug (0 ng/ml), 32 ng/mL, 64 ng/mL).

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