Human stem cells culture

Human embryonic stem cells / Culture as floating aggregratesN. (embryoid bodies), forming / neuroepithelial cells s Culture to form midbrain dopamine neurons ... 

Zambidis E, Oberlin E, Tavian M and Peault B (2006). Blood forming endothelium in human ontogeny lessons from in utero development and embryonic stem cell culture. Trends in Cardiovascular Medicine 16(3) 95 101. 

Progeny of long-term mouse and human CNS stem cell cultures survive transplantation and differentiate into neurons and glia. Borg, L.L., Vescovi, A.L., Blote, K, Kyle, A.L., Hettiaratchi, P., Reynolds, B.A., Mudrick-Donnon, L A. (1996). Abstr SocNeurosci, 22 (1-3) 47. 

Meinel, L., Hofmann, S., Betz, O., Fajardo, R., Merkle, H.P., Langer, R., Evans, C.H., Vunjak-Novakovic, G., and Kaplan, D.L. "Osteogenesis by human mesenchymal stem cells cultured on silk biomaterials Comparison of adenovirus mediated gene transfer and protein delivery of BMP-2". Biomaterials 27(28), 4993-5002 (2006b). 

Human embryonic stem cells were first collected in 1998 by two different research teams. The cells obtained from the inner cell mass of the blastocyst (4- to 5-day embryo) are embryonic stem cells (ESC) in contrast, cells cultured from the primordial germ cells of 5- to 9-week fetuses are embryonic germ cells (EGC). In the laboratory, ES or EG cells can proliferate indefinitely in an undifferentiated state but can also be manipulated to become specialized or partially specialized cell types, a process known as directed differentiation. Both ES and EG cells are pluripotent, meaning they have the potential to develop into more than 200 different known cell types. This class of human stem cells holds the promise of being able to repair or replace cells or tissues that are damaged or destroyed by many of our most devastating diseases and disabilities. 

Neuronal cultures derived from human stem cells can possibly serve as renewable source of normal (non-transformed) cells with the capacity to differentiate into any cell type present in the nervous system. The major advantage of human cell types-based in vitro models is that the results do not require extrapolation from animal data to the human situation. The detailed characterization of human stem/progenitor cell-based assays for neurodevelop-mental toxicity testing is described in Chap. 16 of this book by E. Eritsche. 

Stem cells Culturing human stem cells and differentiating them into dopaminergic neurons for ceU transplantation in PD is a dream that inspires many academic researchers and the biotechnology industry. The idea of creating a rehable and renewable source of uncontaminated, well-defined and characterized cells that could replace degenerated neurons and create the possibility of functional brain repair is indeed intriguing (see also Part 1, Chapters 11, 12 and 15). 

Suzuki, M., Wright, L. S., Marwah, P., Lardy, H. A. and Svendsen, C. N. Mitotic and neurogenic effects of dehydroepiandrosterone (DHEA) on human neural stem cell cultures derived from the fetal cortex. Proc Natl Acad Sci USA 101 (2004) 3202-3207. 

GPI-deficient mammalian cells are viable in tissue culture and many GPI-deficient mutant cell lines have been established. However, GPI deficiency has major consequences at the level of tissues and the whole body. This was revealed in transgenic mouse models in which the PIG-A gene (required for the first step of GPI biosynthesis) was knocked out in specific tissues or in the whole animal. For example, keratinocyte-specific disruption of PIG-A caused abnormal development of skin leading to death of the mutant mice a few days after birth (M. Tarutani, 1997), and disruption of PIG-A in the whole animal resulted in embryos that did not develop beyond day 9 of gestation (M. Nozaki, 1999). A somatic mutation of PIG-A in multipotent hematopoietic human stem cells causes paroxysmal nocturnal hemoglobinuria, an acquired hemolytic disease in humans characterized by abnormal activation of complement on erythrocytes due to a deficiency of GPI-anchored complement regulatory proteins such as decay accelerating factor (N. Inoue, 2003). This disease is characterized by intravascular hemolysis and anemia. 

In the remainder of this section we treat several important aspects of animal cell culture, beginning with a discussion of the significance of mixing and shear stress phenomena in the culture of animal cells. Subsequent sections treat fundamental aspects of the culture of mammahan cells and human stem cells. 

KoUiar P, Kotamraju VR, Hdata ST, Clegg DO, Ruoslahti E. Synthetic surfaces for human embryonic stem cell culture. J Biotechnol 2010 146 143-6. 

Saha K, Mei Y, Reisterer CM, Pyzocha NK, Yang J, Muffat J, et al. Surface-engineered substrates for improved human pluripotent stem cell culture under fuUy defined conditions. Proc Natl Acad Sci 2011 108 18714—9. 

In the United States, the National Institutes of Health supports short-term training courses in human embryonic stem cell culture techniques. These training courses include hands-on experience to improve the knowledge and skills of biomedical researchers to maintain, characterize, and use human embryonic stem cells in basic research studies. The courses are given at various locations throughout the United States. 

G. Meng, S. Liu, X. Li, R. Krawetz, and D.E. Rancourt, Extracellular matrix isolated from foreskin fibroblasts supports long-term xeno-free human embryonic stem cell culture. Stem Cells Develop., 19 (4) 547-556, Apr. 2010. 

K. Rajala, H. Hakala, S. Panula, S. Aivio, H. Pihlajamaki, R. Suuronen, O. Hovatta, and H. Skottman, Testing of nine different xeno-free culture media for human embryonic stem cell cultures. Hum. Reprod., 22 (5) 1231-1238, May. 2007. 

H. Hongisto, S. Vuoristo, A. Mikhailova, R. Suuronen, I. Virtanen, T. Otonkoski, and H. Skottman, Laminin-511 expression is associated with the functionality of feeder cells in human embryonic stem cell culture. Stem Cell Res., 8 (1) 97-108, Jan. 2012. 

M.M. Mahlstedt, D. Anderson, J.S. Sharp, R. McGilvray, M.D. Munoz, E.D. Buttery, M.R. Alexander, ER. Rose, and C. Denning, Maintenance of pluripotency in human embryonic stem cells cultured on a synthetic substrate in conditioned medium, Biotechnol. Bioeng., 105 (1) 130-140, Jan. 2010. 

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