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Three-dimensional co-culture of BM-MSCs and eccrine sweat gland cells in Matrigel promotes transdifferentiation of BM-MSCs

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Abstract

Victims with extensive and deep burns are unable to regenerate eccrine sweat glands. Combining of stem cells and biomimetic ECM to generate cell-based 3D tissues is showing promise for tissue repair and regeneration. We co-cultured BrdU-labeled bone marrow-derived mesenchymal stem cells (BM-MSCs) and eccrine sweat gland cells in Matrigel for 2 weeks in vitro and then evaluated for BM-MSCs differentiation into functional eccrine sweat gland cells by morphological assessment and immunohistochemical double staining for BrdU/pancytokeratin, BrdU/ZO-2, BrdU/E-cadherin, BrdU/desmoglein-2, BrdU/Na+–K+-ATPase α, BrdU/NHE1 and BrdU/CFTR. Cells formed spheroid-like structures in Matrigel, and BrdU-labeled BM-MSCs were involved in the 3D reconstitution of eccrine sweat gland tissues, and the incorporated BM-MSCs expressed an epithelial cell marker (pancytokeratin), epithelial cell junction proteins (ZO-2, E-cadherin and desmoglein-2) and functional proteins of eccrine sweat glands (Na+–K+-ATPase α, NHE1 and CFTR). In conclusion, three-dimensional co-culture of BM-MSCs and eccrine sweat gland cells in Matrigel promotes the transdifferentiation of BM-MSCs into potentially functional eccrine sweat gland cells.

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Abbreviations

3D:

Three-dimensional

AEC:

3-Amino-9-ethylcarbazole

BCIP/NBT:

5-Bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium

BM-MSCs:

Bone marrow-derived mesenchymal stem cells

BPE:

Bovine pituitary extract

CFTR:

Cystic fibrosis transmembrane conductance regulator

DMEM/F12:

Dulbecco’s modified Eagle’s medium/Ham’s F12 (1:1)

ECM:

Extracellular matrix

EGF:

Epidermal growth factor

HE:

Hematoxylin and eosin

FGF:

Fibroblast growth factor

IGF:

Insulin-like growth factor

KSFM:

Keratinocyte serum-free medium

Matrigel:

Matrigel basement membrane matrix

MSCs:

Mesenchymal stem cells

NHE1:

Sodium–hydrogen exchanger 1

PBS:

Phosphate-buffered saline

rhEGF:

Recombinant human epidermal growth factor

TGF-β:

Transforming growth factor beta

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Acknowledgments

The manuscript was supported in part by the National Natural Science Foundation of China (81071551, 81471882) and the Natural Science Foundation of Guangdong Province (2014A030313476).

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Authors and Affiliations

  1. Burn and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, North Dongxia Road, Shantou, 515041, Guangdong Province, China

    Haihong Li, Xuexue Li, Mingjun Zhang, Lu Chen & Shijie Tang

  2. Research Center for Translational Medicine, Shantou University Medical College, North Dongxia Road, Shantou, 515041, Guangdong Province, China

    Haihong Li & Bingna Zhang

  3. Burns Institute, The First Affiliated Hospital, Chinese PLA General Hospital, Trauma Center of Postgraduate Medical School, 51 Fucheng Road, Beijing, 100037, China

    Xiaobing Fu

Authors
  1. Haihong Li
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  2. Xuexue Li
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  3. Mingjun Zhang
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  4. Lu Chen
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  5. Bingna Zhang
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Correspondence to Haihong Li.

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Li, H., Li, X., Zhang, M. et al. Three-dimensional co-culture of BM-MSCs and eccrine sweat gland cells in Matrigel promotes transdifferentiation of BM-MSCs. J Mol Hist 46, 431–438 (2015). https://doi.org/10.1007/s10735-015-9632-5

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  • DOI: https://doi.org/10.1007/s10735-015-9632-5

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