We report on the derivation of a diploid 46(XX) human embryonic stem cell (HESC) line that is homozygous for the common deletion associated with Spinal muscular atrophy type 1 (SMA) from a pathenogenetic embryo. By characterizing the methylation status of three different imprinted loci (MEST, SNRPN and H19), monitoring the expression of two parentally imprinted genes (SNRPN and H19) and carrying out genome-wide SNP analysis, we provide evidence that this cell line was established from the activation of a mutant oocyte by diploidization of the entire genome. Therefore, our SMA parthenogenetic HESC (pHESC) line provides a proof-of-principle for the establishment of diseased HESC lines without the need for gene manipulation. As mutant oocytes are easily obtained and readily available during preimplantation genetic diagnosis (PGD) cycles, this approach should provide a powerful tool for disease modelling and is especially advantageous since it can be used to induce large or complex mutations in HESCs, including gross DNA alterations and chromosomal rearrangements, which are otherwise hard to achieve.

/pdf/establishment-of-homozygote-mutant-human-embryonic-stem-4k46gjwqia.pdf

Establishment of Homozygote Mutant Human Embryonic Stem Cells by Parthenogenesis

Stem cells are immature cells capable of self-renewal and differentiation into mature cells. They are of different types and functionally diverse depending on the site it is produced. A major leap in stem cell culture is seen from using the conventional Mouse Embryo Fibroblast (MEF) with the possibility of problems arising due to xenogenicity to Induced Pluripotent cells (iPS) cells which take the stem cell harvesting to a new level. The usage of stem cells were expanded from human Embryonic stem cell (hESC) to various Mesenchymal Stem cells (MSC), Hematopoietic stem cells (HSC) and humananimal cybrids which showed promise in the treatment of diseases like HIV, Parkinsonism and Alzheimer’s. The stem cell banking made it possible for researchers to further their knowledge and also the patients to utilize the variety of stem cell transplantation obtained from adipose tissue, menstrual blood and dental pulp and thereby not limiting to hESC. In this review we discuss the developing trends in the stem cell culture and also in its banking mainly public with private, considering the facilities available on the storage and accessibility to the researchers and patients. We also discuss about the prerequisites of such storage and the challenges faced by alternative therapeutic possibilities to stem cell therapies. Stem cell banking looks promising amidst its pitfalls however the future trend Review Article British Journal of Medicine & Medical Research, 4(11): 2250-2264, 2014 2251 of stem cell therapy and banking still appears shrouded with unpredictability in the face of newer developments.

Stem Cell and Its Banking: Current Trends and Developments

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Human foreskin fibroblasts: from waste bag to important biomedical applications:

OBJECTIVE To establish a safe, effective, and economic feeder'-free culture system which is suitable for the culture of human parthenogenetic embryonic stem cells (hPESCs) in vitro. METHODS hPESCs were cultured with mTeSR 1 medium (control group) and human foreskin fibroblasts-conditional medium (hFFs-CM) (experimental group). The growth status of hPESCs in both feeder-free culture systems were observed with inverted microscope. Alkaline phosphatase (ALP) analysis and karyotype analysis were used to study the biological characteristics of hPESCs. The expression of hPESCs pluripotent marker Oct-4 was analyzed by RT-PCR. Differentiation experiment in vivo and in vitro was applied to observe the differentiation potential of hPESCs into three germ layers. RESULTS hPESCs had regular morphology with difficulty in differentiation in both culture systems. No obvious difference was observed in morphology and expansion speed of hPESCs between 2 groups. After subcultured for 15 passages in vitro, hPESCs in 2 groups could maintain normal female diploid karyotype 46, XX and pluripotency. The expression of Oct-4 mRNA was positive in 2 groups. hPESCs in 2 groups could form embryonic body in differentiation experiment in vitro and could develop into teratomas containing three germ layers in nude mice. CONCLUSION Feeder-free culture system of hFFs-CM can sustain the growth of hPESCs and keep hPESCs undifferentiated state for long. A feeder-free culture system of hPESCs is successfully established, which can support the growth of hPESCs, reduce the contamination from animals, decrease the cost of culture, and satisfy the clinical large-scale application.

[Establishment of feeder-free culture system of human parthenogenetic embryonic stem cells].

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[Establishment of feeder-free culture system of human parthenogenetic embryonic stem cells].