PMID- 27737727
OWN - NLM
STAT- MEDLINE
DCOM- 20171127
LR  - 20211204
IS  - 1555-3892 (Electronic)
IS  - 0963-6897 (Print)
IS  - 0963-6897 (Linking)
VI  - 26
IP  - 3
DP  - 2017 Mar 13
TI  - Chondrogenic Differentiation of Mesenchymal Stem Cells in Three-Dimensional 
      Chitosan Film Culture.
PG  - 417-427
LID - 10.3727/096368916X693464 [doi]
AB  - Articular cartilage has a very limited capacity for self-repair, and mesenchymal 
      stem cells (MSCs) have the potential to treat cartilage defects and 
      osteoarthritis. However, in-depth mechanistic studies regarding their 
      applications are required. Here we demonstrated the use of chitosan film culture 
      for promoting chondrogenic differentiation of MSCs. We found that MSCs formed 
      spheres 2 days after seeding on dishes coated with chitosan. When MSCs were 
      induced in a chondrogenic induction medium on chitosan films, the size of the 
      spheres continuously increased for up to 21 days. Alcian blue staining and 
      immunohistochemistry demonstrated the expression of chondrogenic proteins, 
      including aggrecan, type II collagen, and type X collagen at 14 and 21 days of 
      differentiation. Importantly, chitosan, with a medium molecular weight (size: 
      190-310 kDa), was more suitable than other sizes for inducing chondrogenic 
      differentiation of MSCs in terms of sphere size and expression of chondrogenic 
      proteins and endochondral markers. We identified that the mechanistic target of 
      rapamycin (mTOR) signaling and its downstream S6 kinase (S6K)/S6 were activated 
      in chitosan film culture compared to that of monolayer culture. The activation of 
      mTOR/S6K was continuously upregulated from days 2 to 7 of differentiation. 
      Furthermore, we found that mTOR/S6K signaling was required for chondrogenic 
      differentiation of MSCs in chitosan film culture through rapamycin treatment and 
      mTOR knockdown. In conclusion, we showed the suitability of chitosan film culture 
      for promoting chondrogenic differentiation of MSCs and its potential in the 
      development of new strategies in cartilage tissue engineering.
FAU - Lu, Tsai-Jung
AU  - Lu TJ
FAU - Chiu, Fang-Yao
AU  - Chiu FY
FAU - Chiu, Hsiao-Ying
AU  - Chiu HY
FAU - Chang, Ming-Chau
AU  - Chang MC
FAU - Hung, Shih-Chieh
AU  - Hung SC
LA  - eng
PT  - Journal Article
DEP - 20161012
PL  - United States
TA  - Cell Transplant
JT  - Cell transplantation
JID - 9208854
RN  - 0 (Aggrecans)
RN  - 0 (Collagen Type II)
RN  - 0 (Collagen Type X)
RN  - 9012-76-4 (Chitosan)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - W36ZG6FT64 (Sirolimus)
SB  - IM
MH  - Aggrecans/metabolism
MH  - Blotting, Western
MH  - Cell Culture Techniques
MH  - Cell Differentiation/drug effects
MH  - Cells, Cultured
MH  - Chitosan/chemistry/*pharmacology
MH  - Chondrogenesis/drug effects
MH  - Collagen Type II/metabolism
MH  - Collagen Type X/metabolism
MH  - Humans
MH  - Immunohistochemistry
MH  - Mesenchymal Stem Cells/*cytology/*drug effects
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sirolimus/pharmacology
MH  - TOR Serine-Threonine Kinases/genetics/metabolism
PMC - PMC5657699
EDAT- 2016/10/16 06:00
MHDA- 2017/11/29 06:00
PMCR- 2017/03/01
CRDT- 2016/10/15 06:00
PHST- 2016/10/16 06:00 [pubmed]
PHST- 2017/11/29 06:00 [medline]
PHST- 2016/10/15 06:00 [entrez]
PHST- 2017/03/01 00:00 [pmc-release]
AID - content-ct-1686_lu_et_al [pii]
AID - 10.3727_096368916X693464 [pii]
AID - 10.3727/096368916X693464 [doi]
PST - ppublish
SO  - Cell Transplant. 2017 Mar 13;26(3):417-427. doi: 10.3727/096368916X693464. Epub 
      2016 Oct 12.