Small molecules facilitate single factor-mediated sweat gland cell reprogramming
Abstract Background Large skin defects severely disrupt the overall skin structure and can irreversibly damage sweat glands (SG), 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...
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| Format: | Article |
| Language: | English |
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BMC
2022-03-01
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| Series: | Military Medical Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40779-022-00372-5 |
| _version_ | 1818773985124417536 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Shuai-Fei Ji |
| collection | DOAJ |
| description | Abstract Background Large skin defects severely disrupt the overall skin structure and can irreversibly damage sweat glands (SG), 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 a 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 EDA overexpression drove 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)% vs. (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. Conclusion 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. |
| first_indexed | 2024-12-18T10:33:56Z |
| format | Article |
| id | doaj.art-d29087a7d70f4647bd58d5d2f918bf2e |
| institution | Directory Open Access Journal |
| issn | 2054-9369 |
| language | English |
| last_indexed | 2024-12-18T10:33:56Z |
| publishDate | 2022-03-01 |
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| series | Military Medical Research |
| spelling | doaj.art-d29087a7d70f4647bd58d5d2f918bf2e2022-12-21T21:10:48ZengBMCMilitary Medical Research2054-93692022-03-019111310.1186/s40779-022-00372-5Small molecules facilitate single factor-mediated sweat gland cell reprogrammingShuai-Fei Ji0Lai-Xian Zhou1Zhi-Feng Sun2Jiang-Bing Xiang3Shao-Yuan Cui4Yan Li5Hua-Ting Chen6Yi-Qiong Liu7Huan-Huan Gao8Xiao-Bing Fu9Xiao-Yan Sun10Research 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 RegenerationResearch 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 RegenerationDepartment of Respiratory, The Second Medical Center, Chinese PLA General HospitalResearch 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 RegenerationDepartment of Nephrology, The First Medical Center, Chinese PLA General Hospital, State Key Laboratory of Kidney DiseasesResearch 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 RegenerationResearch 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 RegenerationResearch 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 RegenerationResearch 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 RegenerationResearch 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 RegenerationResearch 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 RegenerationAbstract Background Large skin defects severely disrupt the overall skin structure and can irreversibly damage sweat glands (SG), 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 a 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 EDA overexpression drove 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)% vs. (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. Conclusion 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.https://doi.org/10.1186/s40779-022-00372-5Direct reprogrammingHuman dermal fibroblastsSweat glandRegeneration |
| spellingShingle | 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 Small molecules facilitate single factor-mediated sweat gland cell reprogramming Military Medical Research Direct reprogramming Human dermal fibroblasts Sweat gland Regeneration |
| title | Small molecules facilitate single factor-mediated sweat gland cell reprogramming |
| title_full | Small molecules facilitate single factor-mediated sweat gland cell reprogramming |
| title_fullStr | Small molecules facilitate single factor-mediated sweat gland cell reprogramming |
| title_full_unstemmed | Small molecules facilitate single factor-mediated sweat gland cell reprogramming |
| title_short | Small molecules facilitate single factor-mediated sweat gland cell reprogramming |
| title_sort | small molecules facilitate single factor mediated sweat gland cell reprogramming |
| topic | Direct reprogramming Human dermal fibroblasts Sweat gland Regeneration |
| url | https://doi.org/10.1186/s40779-022-00372-5 |
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