Morphological evaluation of chondrogenic potency in passaged cell populations
Morphological evaluation of chondrogenic potency in passaged cell populations
复制标题
DOI：
10.1016/j.jbiosc.2008.12.018
复制
发表时间：
2009-05-01
影响因子：
2.8
通讯作者：
Taya, Masahito
中科院分区：
文献类型：
--
作者：
Kino-oka, Masahiro;Maeda, Yoshikatsu;Taya, Masahito
本研究描述了在细胞扩增过程中，通过在不同水平的群体doubledore（PD）的T-烧瓶中连续传代培养兔软骨细胞的软骨形成能力的形态学评估与传统的聚苯乙烯表面。将传代群体接种在高密度胶原表面（CL表面）和胶原凝胶（CL凝胶）支架上，以分别评估软骨细胞的平面和空间形态，以及I型和II型胶原mRNA的基因表达。平面形态估计是基于在CL表面上孵育1天后不同PD值下软骨细胞的圆度（R-c）。随着PD值的增加，R-c > 0.9（f（R））的圆形细胞的频率降低，同时I型胶原mRNA水平增加。在PD = 17.8时，频率达到f（R）= 0.12，小于PD = 0时的六分之一。通过在接种后4天测定的球形度（S-c）方面估计空间形态，发现包埋在CL凝胶中的传代软骨细胞存在类似趋势。随着PD值的增加，S-c > 0.9（fs）的球形细胞的频率降低，I型胶原mRNA表达增加，在PD = 17.8时f（s）= 0.28，不到PD = 0时的四分之一。从这些结果中，CL表面和CL凝胶中的细胞形态被提出作为了解软骨形成潜力的指标，这些潜力涉及细胞扩增过程中群体中的表型和分化状态，最终导致组织工程软骨的质量控制。(c)2009年，生物技术学会，日本。All rights reserved.
The present study describes the morphological assessment of chondrogenic potency during a cell expanding process through serial subculturing of rabbit chondrocytes at different levels of population doublings; (PD) in a T-flask with a conventional polystyrene surface. The passaged populations were seeded on a high-density collagen surface (CL surface) and in a collagen gel (CL gel) scaffold to evaluate the planar and spatial morphologies of the chondrocytes, respectively, as well as the gene expressions of mRNA for collagen types I and II. The planar morphological estimation was based on roundness (R-c) of chondrocyte cells at different PD values after I day incubation on the CL surface. The frequency of round-shaped cells with R-c > 0.9 (f(R)) decreased with increasing PD values, accompanied by an increase in collagen type I mRNA level. At PD = 17.8, the frequency reached f(R) = 0.12, which was less than one-sixth of that at PD = 0. A similar trend was found with respect to the passaged chondrocytes embedded in the CL gels by estimating the spatial morphology in terms of sphericity (S-c) determined 4 days after seeding. With an increase in PD value, the frequency in spherical-shaped cells with S-c > 0.9 (fs) decreased and the mRNA expression of collagen type I increased, giving f(s) = 0.28 at PD = 17.8 which was less than a quarter of that at PD = 0. From these results, the cell morphologies on the CL surface and in the CL gel were proposed as indicators for understanding chondrogenic potentials concerning the phenotypes and differentiated states in the population during cell expansion, ultimately leading to quality control of tissue-engineered cartilage. (c) 2009, The Society for Biotechnology, Japan. All rights reserved.