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Keratinocyte growth factor and thiazolidinediones and linolenic acid differentiate characterized mammary fat pad adipose stem cells isolated from prepubertal Korean black goat to epithelial and adipogenic lineage

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Abstract

The study was conducted to know and investigate the mechanism involved during mesenchymal to epithelial transition to unravel questions related to mammary gland development in prepubertal Korean black goat. We, therefore, biopsied mammary fat pad and isolated adipose cells and characterized with stemness factors (CD34, CD13, CD44, CD106, and vimentin) immunologically and through their genetic expression. Furthermore, characterized cells were differentiated to adipogenic (thiazolidinediones and α-linolenic acid) and epithelial (keratinocyte growth factor) lineages. Thiazolidinediones/or in combination with α-linolenic acid demonstrated significant upregulation of adipo-Q, PPAR-γ, CEBP-α, LPL, and resistin. Adipose stem cells in induction mixture (5 μg/ml insulin, 1 μg/ml hydrocortisone, and 10 ng/ml epidermal growth factor) and subsequent treatment with 10 ng/ml keratinocyte growth factor revealed their trans-differentiating ability to epithelial lineage. From 2 d onwards, the cells under keratinocyte growth factor influenced cells to assume rectangular (2–4 d) to cuboidal (8–10 d) shapes. Ayoub–Shklar stain developed brownish-red pigment in the transformed cells. Though, expressions of K8 and K18 were noted to be highly significant (p < 0.01) but expressions of epithelial membrane antigens and epithelial specific antigens were also significant (p < 0.05) compared to 0 d. Conclusively, epithelial transformations of mammary adipose stem cells would add up knowledge to develop therapeutic regimen to deal with mammary tissue injury and diseases.

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Acknowledgments

The research was fully supported and facilitated from National Research Foundation of Korea (grant no.120130344) and was partly supported by research grant from Kangwon national University (grant no. 120100910) and (grant no. 120131475).

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Authors and Affiliations

  1. Department of Animal Biotechnology, College of Animal Life Sciences, Kangwon National University, Chuncheon, Republic of Korea

    A. M. M. T Reza, S. Shiwani, N. K. Singh, J. D. Lohakare & S. J. Lee

  2. Department of Animal Biotechnology, College of Applied Life Sciences, Jeju National University, Jeju, Republic of Korea

    D. K. Jeong

  3. WCU Biomodulation Major, Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea

    J. Y. Han & D. Rengaraj

  4. College of Veterinary Medicine, Kangwon National University, Chuncheon, Republic of Korea

    B. W. Lee

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  1. A. M. M. T Reza
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Reza, A.M.M.T., Shiwani, S., Singh, N.K. et al. Keratinocyte growth factor and thiazolidinediones and linolenic acid differentiate characterized mammary fat pad adipose stem cells isolated from prepubertal Korean black goat to epithelial and adipogenic lineage. In Vitro Cell.Dev.Biol.-Animal 50, 194–206 (2014). https://doi.org/10.1007/s11626-013-9690-5

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