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Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

Restu Syamsul Hadi, Indra Kusuma, Yurika Sandra
Submission date: Monday, 30 November 2015
Published date: Wednesday, 02 December 2015
DOI: http://dx.doi.org/10.18051/UnivMed.2014.v33.91-99

Abstract


BACKGROUND

Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC).

METHODS

In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4) expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1) proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test.

RESULTS

Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05). Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity

CONCLUSIONS

Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.


Keywords


Fibroblast; PBMC; co-culture; foreskin

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References


Li L, Fukunaga-Kalabis M, Yu H, Xu X, Kong J, Lee JT, et al. Human dermal stem cells differentiate into functional epidermal melanocytes. J Cell Sci 2010;123:853–60.

Bi D, Chen FG, Zhang WJ, Zhou GD, Cui L, Liu W, et al. Differentiation of human multipotent dermal fibroblasts into islet-like cell clusters. BMC Cell Biol 2010;11:46.

Crigler L, Kazhanie A, Yoon TJ, Zakhari J, Anders J, Taylor B, et al. Isolation of a mesenchymal cell population from murine dermis that contains progenitors of multiple cell lineages. FASEB J 2007;21:2050–63.

Shim JH, Park JY, Lee MG, Kang HH, Lee TR, Shin DW. Human dermal stem/progenitor cell-derived conditioned medium ameliorates ultraviolet Ainduced damage of normal human dermal fibroblasts. PloS One 2013;8:e67604.

Crespo AV, Awe JP, Reijo Pera R, Byrne JA. Human skin cells that express stage-specific embryonic antigen 3 associate with dermal tissue regeneration. Bio Res 2012;1:25–33.

Bakker K, Schaper N. The development of global consensus guidelines on the management and prevention of the diabetic foot 2011. Diabetes Met Res Rev 2012;28:116–8.

Ding W, Stohl LL, Wagner JA, Granstein RD. Calcitonin gene-related peptide biases Langerhans cells toward Th2-type immunity. J Immunol 2008;181:6020–6.

Mukonoweshuro B, Brown CJ, Fisher J, Ingham E. Immunogenicity of undifferentiated and differentiated allogeneic mouse mesenchymal stem cells. J Tissue Eng 2014;5:2041731414534255. doi:10.1177/2041731414534255.

Mizukami Y, Abe T, Shibata H, Makimura Y, Fujishiro SH, Yanase K, et al. MHC-matched induced pluripotent stem cells can attenuate cellular and humoral immune responses but are still susceptible to innate immunity inpigs. PloS One 2014;9:e98319.

Hargreave T. Male circumcision: towards a World Health Organisation normative practice in resource limited settings. Asian J Androl 2010;12:628–38.

Kusuma I, Hadi RS. Geraniin supplementation increases human keratinocyte proliferation in serum-free culture. Univ Med 2013;32:3-10.

Djuwantono T, Wirakusumah FF, Achmad TH, Sandra F, Halim D, Faried A. A comparison of cryopreservation methods: slow-cooling vs. rapid-cooling based on cell viability, oxidative stress, apoptosis, and CD34+ enumeration of human umbilical cord blood mononucleated cells. BMC Research Notes 2011;4:371.

Coolen NA, Schouten KC, Middelkoop E, Ulrich MM. Comparison between human fetal and adult skin. Arch Dermatol Res 2010;302:47–55.

Pereira C, Gold W, Herndon D. Review paper: burn coverage technologies: current concepts and future directions. J Biomater Applic 2007;22:101–21.

Auger F, Lacroix D, Germain L. Skin substitutes and wound healing. Skin Pharmacol Physiol 2009; 22:94–102.

Pereyra-Bonnet F, Gimeno ML, Argumedo NR, Ielpi M, Cardozo JA, Gimenez, et al. Skin fibroblasts from patients with type 1 diabetes (T1D) can be chemically transdifferentiated into insulin-expressing clusters: atransgene-free approach. PloS One 2014;9:e100369.

Simeonov KP, Uppal H. Direct reprogramming of human fibroblasts to hepatocyte-like cells by synthetic modified mRNAs. PloS One 2014;9: e100134.

Durruthy-Durruthy J, Briggs SF, Awe J, Ramathal CY, Kayumbaran S, Lee PC, et al. Rapid and efficient conversion of integration-free human induced pluripotent stem cells to GMP-grade culture conditions. PloS One 2014;9:e94231.

Curnow SJ, Fairclough M, Schmutz C, Kissane S, Denniston AKO, Nash K, et al. Distinct types of fibrocyte can differentiate from mononuclear cells in the presence and absence of serum. PLoS One 2010;5:e9730.

Pilling D, Vakil V, Gomer RH. Improved serum-free culture conditions for the differentiation of human and murine fibrocytes. J Immunol Methods 2009; 351:62–70.


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