Chemically‐Defined Culture of Human Embryonic Stem Cells

Clark, Julie; Ding, Sheng

doi:10.1002/9780470511619.ch6

Cited by 1 publication

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“…Although the number of cells expressing MBP almost duplicated from 3 to 9 DIV, we assume that our culture medium is not appropriate to further increment or at least to maintain constant the number of MBP positive cells. The bovine serum albumin present in our differentiating protocol, as well as the one in the B-27 supplement, by containing some bone morphogenic protein-like activity (Clark and Ding, 2007), may have contributed to inhibit further oligodendrocyte precursor cell maturation (See et al, 2004). Nevertheless, if desirable, both NG2 and MBP enriched cultures may be obtained by magnetic cell sorting (Cizkova et al, 2009).…”

Section: Resultsmentioning

confidence: 99%

Directing mouse embryonic neurosphere differentiation toward an enriched neuronal population

Torrado

Gomes

Santos

et al. 2014

Intl J of Devlp Neuroscience

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Neural stem cells (NSC) are self-renewing multipotent cells that have emerged as a powerful tool to repair the injured brain. These cells can be cultured as neurospheres, which are floating aggregates of neural stem/progenitor cells (NSPCs). Despite their high clonal expansion capacity, it has been suggested that in neurospheres, only a small percentage of cells are capable of proliferation and that this system is not efficient in terms of neurogenic competence. Thus, our aim was to develop a neurosphere culture method with a highly proliferative stem/progenitor cell population and particularly with a prominent neurogenic potential, surpassing some of the claimed weaknesses of the neurosphere assay. In our model, mouse neurospheres were harvested from neural tissue at E15 and after only 4 days in vitro (DIV), we have achieved highly proliferative primary neurospheres (81% Sox2 and 76% Ki67 positive cells) and a rather low number of cells expressing glial and neuronal markers (∼10%). After inducing differentiation, we have attained an enriched neuronal population (45% β-III-tubulin positive cells at 15 DIV). Using a simple methodology, we have developed a NSPC model that can provide a valuable source of neuronal precursors, thus offering a potential starting point for cell replacement therapies following CNS injury.

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Section: Resultsmentioning

confidence: 99%