Effects of type IV collagen and laminin on the cryopreservation of human embryonic stem cells.
Previous reports have indicated that extracellular matrices (ECMs) affect the developmental fate of human embryonic stem cells (hESCs). Specially, type IV collagen and laminin, which belong to a group of macromolecular proteins with a substantial proportion of ECMs, are known to influence the proliferation and differentiation of hES cells. In this study, we evaluated the effects of type IV collagen and laminin in freezing medium on the survival and differentiation rates of hES cells after slow freezing and rapid thawing. The addition of type IV collagen (1 microg/ml) to the freezing medium significantly increased the survival rate of hES cells after thawing compared with that of a control group. The spontaneous differentiation rates of groups treated with type IV collagen (1 microg/ml) or laminin (1 microg/ml) were significantly lower than those of the control group. Frozen-thawed hES cells have currently been cultured for more than 70 passages and retain key properties of hES cells such as morphological characteristics, normal karyotype, marker expression (alkaline phosphatase, SSEA-1, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, Rex-1, and Oct-4), basement membrane-related gene expression, and the potential to differentiate into derivatives of all three germ layers. This new slow freezing method by ECM treatment is a reliable and effective cryopreservation method for pluripotent hES cells.
Kim
Sun Jong
SJ
Division of Stem Cell Biology, Medical Research Center, MizMedi Hospital, Kangseo-ku, Seoul 157-280, Korea.
Park
Jong Hyuk
JH
Lee
Jeoung Eun
JE
Kim
Jin Mee
JM
Lee
Jung Bok
JB
Moon
Shin Yong
SY
Roh
Sung Il
SI
Kim
Chul Geun
CG
Yoon
Hyun Soo
HS
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D000954:Antigens, Surface; D015295:Antigens, Tumor-Associated, Carbohydrate; D001485:Basement Membrane / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival; D017690:Cell Transplantation; D024141:Collagen Type IV / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D004622:Embryo, Mammalian / Q000166:cytology*; D005434:Flow Cytometry; D015870:Gene Expression; D006023:Glycoproteins / Q000096:biosynthesis; D006028:Glycosphingolipids / Q000096:biosynthesis; D020662:Guanine Nucleotide Exchange Factors / Q000096:biosynthesis; D006801:Humans; D007621:Karyotyping; D007797:Laminin / Q000378:metabolism / Q000502:physiology*; D016256:Lewis X Antigen / Q000096:biosynthesis; D051379:Mice; D016513:Mice, SCID; D050814:Octamer Transcription Factor-3; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055746:Stage-Specific Embryonic Antigens; D013234:Stem Cells / Q000166:cytology*; D013696:Temperature; D013724:Teratoma / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis
D000954:Antigens, Surface; D015295:Antigens, Tumor-Associated, Carbohydrate; D024141:Collagen Type IV; D003470:Culture Media; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D006028:Glycosphingolipids; D020662:Guanine Nucleotide Exchange Factors; D007797:Laminin; D016256:Lewis X Antigen; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C453280:PREX1 protein, human; C494174:Pou5f1 protein, mouse; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; C107064:TRA-1-60 antigen, human; D014157:Transcription Factors; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4; D000469:Alkaline Phosphatase