Embryoid bodies formation and differentiation from mouse embryonic stem cells in collagen/Matrigel scaffolds.
Embryonic stem (ES) cells have the potential to develop into any type of tissue and are considered as a promising source of seeding cells for tissue engineering and transplantation therapy. The main catalyst for ES cells differentiation is the growth into embryoid bodies (EBs), which are utilized widely as the trigger of in vitro differentiation. In this study, a novel method for generating EBs from mouse ES cells through culture in collagen/Matrigel scaffolds was successfully established. When single ES cells were seeded in three dimensional collagen/Matrigel scaffolds, they grew into aggregates gradually and formed simple EBs with circular structures. After 7 days' culture, they formed into cystic EBs that would eventually differentiate into the three embryonic germ layers. Evaluation of the EBs in terms of morphology and potential to differentiate indicated that they were typical in structure and could generate various cell types; they were also able to form into tissue-like structures. Moreover, with introduction of ascorbic acid, ES cells differentiated into cardiomyocytes efficiently and started contracting synchronously at day 19. The results demonstrated that collagen/Matrigel scaffolds supported EBs formation and their subsequent differentiation in a single three dimensional environment.
Zhou
Jin
J
Department of Tissue Engineering, Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing 100850, China.
Zhang
Ye
Y
Lin
Qiuxia
Q
Liu
Zhiqiang
Z
Wang
Haibin
H
Duan
Cuimi
C
Wang
Yanmeng
Y
Hao
Tong
T
Wu
Kuiwu
K
Wang
Changyong
C
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D003094:Collagen; D004338:Drug Combinations; D004622:Embryo, Mammalian; D058732:Embryoid Bodies / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation; D005855:Germ Layers; D007797:Laminin; D051379:Mice; D011509:Proteoglycans; D013234:Stem Cells
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen