PMID- 33562279
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210303
IS  - 2077-0383 (Print)
IS  - 2077-0383 (Electronic)
IS  - 2077-0383 (Linking)
VI  - 10
IP  - 4
DP  - 2021 Feb 7
TI  - Endothelial Nitric Oxide Synthase Knockdown in Human Stem Cells Impacts 
      Mitochondrial Biogenesis and Adipogenesis: Live-Cell Real-Time Fluorescence 
      Imaging.
LID - 10.3390/jcm10040631 [doi]
LID - 631
AB  - We carried out live-cell real-time fluorescence imaging to follow the effects of 
      genetic (siRNA) knockdown (KD) of endothelial nitric oxide synthase (eNOS) on 
      mitochondrial biogenesis and adipogenesis in human mesenchymal stem cells 
      (hMSCs). We report here that eNOS KD in hMSCs blocks mitochondrial biogenesis and 
      adipogenesis. The transfer of mitochondria from normal hMSCs to eNOS-deficient 
      hMSCs restores adipogenesis. Furthermore, cell-free mitochondria purified from 
      normal hMSCs also restores adipogenesis in eNOS-deficient cells. Thus, eNOS and 
      NO signaling are essential for mitochondrial biogenesis, and mitochondrial 
      activity is indispensable for adipogenesis in hMSC differentiation. We mapped the 
      path and identified the mechanisms of mitochondrial transfer. We captured 
      real-time images of differentiated mature adipocytes in mitosis and replication. 
      These results reveal that human stem cell-differentiated fat cells are capable of 
      replication. This new finding offers novel insights into our understanding of fat 
      cell expansion and the development of obesity. Real-time imaging in live cells 
      allows synchronized investigation of mitochondrial biogenesis and adipogenesis in 
      stem cell differentiation without reducing living cells to nonliving samples for 
      functional analysis. Live-cell real-time imaging can thus be a faithful and 
      immediate tool for molecular diagnostic medicine. Furthermore, our results 
      suggest that mitochondrial remodeling can be a useful approach in treating 
      adiposity, diabetes, and abnormalities in energy metabolism and vascular 
      signaling.
FAU - Lee-Huang, Sylvia
AU  - Lee-Huang S
AD  - Department of Biochemistry and Molecular Pharmacology, NYU Grossman School of 
      Medicine, New York University, New York, NY 10016, USA.
FAU - Huang, Philip Lin
AU  - Huang PL
AD  - Department of Medicine, Harvard Medical School and Massachusetts General 
      Hospital, Boston, MA 02114, USA.
FAU - Huang, Paul Lee
AU  - Huang PL
AD  - Department of Medicine, Harvard Medical School and Massachusetts General 
      Hospital, Boston, MA 02114, USA.
LA  - eng
GR  - NF-43/Blackout Funds/
PT  - Journal Article
DEP - 20210207
PL  - Switzerland
TA  - J Clin Med
JT  - Journal of clinical medicine
JID - 101606588
PMC - PMC7914526
OTO - NOTNLM
OT  - adipogenesis
OT  - endothelial nitric oxide synthase (eNOS)
OT  - human mesenchymal stem cell (hMSC)
OT  - live-cell real-time fluorescence imaging
OT  - mitochondrial biogenesis
OT  - mitochondrial remodeling
OT  - molecular diagnostic medicine
OT  - nitric oxide (NO)
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses, or interpretation of data; in 
      the writing of the manuscript; or in the decision to publish the results.
EDAT- 2021/02/11 06:00
MHDA- 2021/02/11 06:01
PMCR- 2021/02/07
CRDT- 2021/02/10 01:01
PHST- 2020/12/20 00:00 [received]
PHST- 2021/01/25 00:00 [revised]
PHST- 2021/01/27 00:00 [accepted]
PHST- 2021/02/10 01:01 [entrez]
PHST- 2021/02/11 06:00 [pubmed]
PHST- 2021/02/11 06:01 [medline]
PHST- 2021/02/07 00:00 [pmc-release]
AID - jcm10040631 [pii]
AID - jcm-10-00631 [pii]
AID - 10.3390/jcm10040631 [doi]
PST - epublish
SO  - J Clin Med. 2021 Feb 7;10(4):631. doi: 10.3390/jcm10040631.