PMID- 33417759
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210114
IS  - 2373-9878 (Electronic)
IS  - 2373-9878 (Linking)
VI  - 5
IP  - 8
DP  - 2019 Aug 12
TI  - Remnant Effects of Culture Density on Cell Chirality After Reseeding.
PG  - 3944-3953
LID - 10.1021/acsbiomaterials.8b01364 [doi]
AB  - Proper muscle function requires specific orientation of myotubes. Cell chirality, 
      a mechanical behavior of cells, may participate in myogenesis and give rise to 
      left-right (LR) orientation of muscle tissue. Thus, it is essential to understand 
      the factors effecting the cell chirality. Here, using C2C12 cells as a model 
      system, we report that prior culture condition with high/low density can create 
      remnant effects on cell chirality after reseeding. C2C12 myoblasts were first 
      conditioned by a series of subcultures with plating density at 2200 cells/cm(2) 
      (low density) or 22 000 cells/cm(2) (high density). After reseeding on 
      micropatterned stripes fabricated on glass or polydimethylsiloxane (PDMS) 
      substrates, we found that the cells after low-density cultures exhibited a 
      reduced cell aspect ratio and intercellular alignment, leading to an attenuated 
      chiral orientation only appearing on glass substrate. In contrast, chiral 
      orientation was observed in cells after high-density culture on both substrates. 
      By comparing it to the original cells without being subcultured with high/low 
      density, we found that the series of low-density cultures disorganized the 
      formation of actin rings in single cells, which is an essential structure for 
      cell chirality. Moreover, by using high-density culture supplemented with 
      inhibitors of actin polymerization, the effect of low-density cultures was 
      recaptured, suggesting that the series of subcultures with high/low density may 
      be an in vitro aging process that modifies the actin cytoskeleton, causing a 
      remnant attenuation of cell chirality even after trypsin digestion and reseeding. 
      Together, our result suggests a mechanistic insight of how cytoskeletal 
      structures "memorize" the previous experience through modification of the actin 
      filament, opening up new possibilities for morphogenesis and mechanobiology.
FAU - Kwong, Hoi Kwan
AU  - Kwong HK
FAU - Huang, Yaozhun
AU  - Huang Y
FAU - Bao, Yuanye
AU  - Bao Y
FAU - Lam, Miu Ling
AU  - Lam ML
AD  - CityU Shenzhen Research Institute, Shenzhen 518057, China.
FAU - Chen, Ting-Hsuan
AU  - Chen TH
AUID- ORCID: 0000-0003-4517-7750
AD  - CityU Shenzhen Research Institute, Shenzhen 518057, China.
AD  - State Key Laboratory of Molecular Engineering of Polymers, Fudan University, 
      Shanghai 200086, China.
LA  - eng
PT  - Journal Article
DEP - 20190604
PL  - United States
TA  - ACS Biomater Sci Eng
JT  - ACS biomaterials science & engineering
JID - 101654670
SB  - IM
OTO - NOTNLM
OT  - cell chirality
OT  - cell density
OT  - cell orientation
OT  - left-right asymmetry
OT  - micropatterning
EDAT- 2019/08/12 00:00
MHDA- 2019/08/12 00:01
CRDT- 2021/01/08 20:11
PHST- 2021/01/08 20:11 [entrez]
PHST- 2019/08/12 00:00 [pubmed]
PHST- 2019/08/12 00:01 [medline]
AID - 10.1021/acsbiomaterials.8b01364 [doi]
PST - ppublish
SO  - ACS Biomater Sci Eng. 2019 Aug 12;5(8):3944-3953. doi: 
      10.1021/acsbiomaterials.8b01364. Epub 2019 Jun 4.