Small molecules facilitate single factor-mediated sweat gland cell reprogramming

Shuai-Fei Ji; Lai-Xian Zhou; Zhi-Feng Sun; Jiang-Bing Xiang; Shao-Yuan Cui; Yan Li; Hua-Ting Chen; Yi-Qiong Liu; Huan-Huan Gao; Xiao-Bing Fu; Xiao-Yan Sun

doi:10.1186/s40779-022-00372-5

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Small molecules facilitate single factor-mediated sweat gland cell reprogramming

RESEARCH | Updated：2023-03-20

- Small molecules facilitate single factor-mediated sweat gland cell reprogramming
- Military Medical Research Vol. 9, Issue 6, Pages: 655-667(2022)
- Affiliations：
  
  1.Research Center for Tissue Repair and Regeneration Affiliated To Medical Innovation Research Department and 4th Medical Center, PLA General Hospital and PLA Medical College; PLA Key Laboratory of Tissue Repair and Regenerative Medicine and Beijing Key Research Laboratory of Skin Injury, Repair and Regeneration, 28 Fu Xing Road, Beijing 100853, China
  2.Research Unit of Trauma Care, Tissue Repair and Regeneration, Chinese Academy of Medical Sciences, 2019RU051, Beijing 100048, China
  3.Department of Respiratory, the Second Medical Center, Chinese PLA General Hospital, Beijing 100036, China
  4.Bioengineering College of Chongqing University, Chongqing 400044, China
  5.Department of Nephrology, the First Medical Center, Chinese PLA General Hospital, State Key Laboratory of Kidney Diseases, Beijing 100048, China
- Author bio：
  
  * fuxiaobing@vip.sina.com;
  yanzisun1979@sina.com
- Funds：
- DOI：10.1186/s40779-022-00372-5
  CLC：
- Published：2022-12
- Accepted：
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Ji SF, Zhou LX, Sun ZF, Xiang JB, Cui SY, Li Y, et al. Small molecules facilitate single factor-mediated sweat gland cell reprogramming. Mil Med Res. 2022;9(1):13.
DOI：

Ji SF, Zhou LX, Sun ZF, Xiang JB, Cui SY, Li Y, et al. Small molecules facilitate single factor-mediated sweat gland cell reprogramming. Mil Med Res. 2022;9(1):13. DOI： 10.1186/s40779-022-00372-5.

摘要

Abstract

Background:

Large skin defects severely disrupt the overall skin structure and can irreversibly damage sweat glands (SGs)

thus impairing the skin’s physiological function. This study aims to develop a stepwise reprogramming strategy to convert fibroblasts into SG lineages

which may provide a promising method to obtain desirable cell types for the functional repair and regeneration of damaged skin.

Methods:

The expression of the SG markers cytokeratin 5 (CK5)

cytokeratin 10 (CK10)

cytokeratin 18 (CK18)

carcino-embryonic antigen (CEA)

aquaporin 5 (AQP5) and α-smooth muscle actin (α-SMA) was assessed with quantitative PCR (qPCR)

immunofluorescence and flow cytometry. Calcium activity analysis was conducted to test the function of induced SG-like cells (iSGCs). Mouse xenograft models were also used to evaluate the

in vivo

regeneration of iSGCs. BALB/c nude mice were randomly divided into normal group

SGM treatment group and iSGC transplantation group. Immunocytochemical analyses and starch-iodine sweat tests were used to confirm the

in vivo

regeneration of iSGCs.

Results:

Ectodermal dysplasia antigen (EDA) overexpression drove human dermal fibroblast (HDF) conversion into iSGCs in SG culture medium (SGM). qPCR indicated significantly increased mRNA levels of the SG markers CK5

CK18 and CEA in iSGCs

and flow cytometry data demonstrated (4.18±0.04)% of iSGCs were CK5 positive and (4.36±0.25)% of iSGCs were CK18 positive. The addition of chemical cocktails greatly accelerated the SG fate program. qPCR results revealed significantly increased mRNA expression of CK5

CK18 and CEA in iSGCs

as well as activation of the duct marker CK10 and luminal functional marker AQP5. Flow cytometry indicated

after the treatment of chemical cocktails

(23.05±2.49)% of iSGCs expressed CK5

and (55.79±3.18)% of iSGCs expressed CK18

respectively. Calcium activity analysis indicated that the reactivity of iSGCs to acetylcholine was close to that of primary SG cells [(60.79±7.71)%

. (70.59±0.34)%

ns].

In vivo

transplantation experiments showed approximately (5.2±1.1)% of the mice were sweat test positive

and the histological analysis results indicated that regenerated SG structures were present in iSGCs-treated mice.

Conclusions:

We developed a SG reprogramming strategy to generate functional iSGCs from HDFs by using the single factor EDA in combination with SGM and small molecules. The generation of iSGCs has important implications for future

in situ

skin regeneration with SG restoration.

关键词

Keywords

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