Role of insulin-transferrin-selenium in auricular chondrocyte proliferation and engineered cartilage formation in vitro.
The goal of this study is to determine the effects of Insulin-Transferrin-Selenium (ITS) on proliferation of auricular chondrocytes and formation of engineered cartilage in vitro. Pig auricular monolayer chondrocytes and chondrocyte pellets were cultured in media containing 1% ITS at different concentrations of fetal bovine serum (FBS, 10%, 6%, 2%, 0%), or 10% FBS alone as a control for four weeks. Parameters including cell proliferation in monolayer, wet weight, collagen type I/II/X (Col I, II, X) and glycosaminoglycan (GAG) expression, GAG content of pellets and gene expression associated with cartilage formation/dedifferentiation (lost cartilage phenotype)/hypertrophy within the chondrocyte pellets were assessed. The results showed that chondrocytes proliferation rates increased when FBS concentrations increased (2%, 6%, 10% FBS) in ITS supplemented groups. In addition, 1% ITS plus 10% FBS significantly promoted cell proliferation than 10% FBS alone. No chondrocytes grew in ITS alone medium. 1% ITS plus 10% FBS enhanced cartilage formation in terms of size, wet weight, cartilage specific matrices, and homogeneity, compared to 10% FBS alone group. Furthermore, ITS prevented engineered cartilage from dedifferentiation (i.e., higher index of Col II/Col I mRNA expression and expression of aggrecan) and hypertrophy (i.e., lower mRNA expression of Col X and MMP13). In conclusion, our results indicated that ITS efficiently enhanced auricular chondrocytes proliferation, retained chondrogenic phenotypes, and promoted engineered cartilage formation when combined with FBS, which is potentially used as key supplementation in auricular chondrocytes and engineered cartilage culture.
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. xliu0810@gmail.com.
Xia
X
Liu
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. B2008183@gmail.com.
Jinchun
J
Liu
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. eilain1983@gmail.com.
Ning
N
Kang
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. yanli102894@gmail.com.
Li
L
Yan
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. liousy6529@gmail.com.
Qian
Q
Wang
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. xiaoxin8349@gmail.com.
Xin
X
Fu
Wake Forest Institute of Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA. yzhang@wakehealth.edu.
Yuanyuan
Y
Zhang
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. xiaoran@pumc.edu.cn.
Ran
R
Xiao
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100144, China. yilincao.rc@gmail.com.
Yilin
Y
Cao
D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004425:Ear Cartilage / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D006025:Glycosaminoglycans / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012643:Selenium / Q000494:pharmacology*; D013552:Swine; D023822:Tissue Engineering; D014168:Transferrin / Q000494:pharmacology*
D053674:Aggrecans; D016895:Culture Media, Serum-Free; D006025:Glycosaminoglycans; D007328:Insulin; D012333:RNA, Messenger; D014168:Transferrin; D003094:Collagen; D012643:Selenium