Human-serum matrix supports undifferentiated growth of human embryonic stem cells.
One of the most frequently used matrices for feeder-free growth of undifferentiated human embryonic stem cells (hESCs) is Matrigel, which supports attachment and growth of undifferentiated hESCs in the presence of mouse embryonic fibroblast-conditioned medium. Unfortunately, application of Matrigel or medium conditioned by mouse embryonic feeder cells is not ideal for potential medical application of hESCs because xenogeneic pathogens can be transmitted through culture conditions. We demonstrate here that human serum as matrix and medium conditioned by differentiated hESCs reduce exposure of hESCs to animal ingredients and provide a safer direction toward completely animal-free conditions for application, handling, and understanding of hESC biology. At the same time, hESCs grown under these conditions maintain all hESC features after prolonged culture, including the developmental potential to differentiate into representative tissues of all three embryonic germ layers, unlimited and undifferentiated proliferative ability, and maintenance of normal karyotype.
Stojkovic
Petra
P
Centre for Stem Cell Biology and Developmental Genetics, Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne, UK.
Lako
Majlinda
M
Przyborski
Stefan
S
Stewart
Rebecca
R
Armstrong
Lyle
L
Evans
Jerome
J
Zhang
Xin
X
Stojkovic
Miodrag
M
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D003094:Collagen / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D017931:DNA Primers / Q000737:chemistry; D004338:Drug Combinations; D004622:Embryo, Mammalian / Q000166:cytology*; D006801:Humans; D007621:Karyotyping / Q000379:methods; D007797:Laminin / Q000494:pharmacology; D008297:Male; D051379:Mice; D016513:Mice, SCID; D008855:Microscopy, Electron, Scanning; D011509:Proteoglycans / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology*; D013737:Testis / Q000378:metabolism
D017077:Culture Media, Conditioned; D017931:DNA Primers; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen