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Journal of Molecular Histology

, Volume 46, Issue 4–5, pp 431–438 | Cite as

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

  • Haihong LiEmail author
  • Xuexue Li
  • Mingjun Zhang
  • Lu Chen
  • Bingna Zhang
  • Shijie Tang
  • Xiaobing Fu
Original Paper

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.

Keywords

Mesenchymal stem cells Eccrine sweat gland cells Matrigel Three-dimensional co-culture Transdifferentiation 

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

Notes

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).

Compliance with ethical standards

Conflict of interest

We declare we have no competing financial, personal or other relationships with other people or organizations.

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Haihong Li
    • 1
    • 2
    Email author
  • Xuexue Li
    • 1
  • Mingjun Zhang
    • 1
  • Lu Chen
    • 1
  • Bingna Zhang
    • 2
  • Shijie Tang
    • 1
  • Xiaobing Fu
    • 3
  1. 1.Burn and Plastic Surgery, The Second Affiliated HospitalShantou University Medical CollegeShantouChina
  2. 2.Research Center for Translational MedicineShantou University Medical CollegeShantouChina
  3. 3.Burns Institute, The First Affiliated Hospital, Chinese PLA General HospitalTrauma Center of Postgraduate Medical SchoolBeijingChina

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