Amniotic Fluid: New Source for Stem Cell Research

Introduction:

In the research paper, "Oct-4-expressing cells in human amniotic fluid: a new source for stem cell research?" (link is located at the bottom of this article),  scientists try to use amniotic fluid of a pregnant woman as a new source of stem cell research while avoiding the ethical concerns that usually overwhelms such studies. (A)

This study investigated whether cells that express transcription factor Oct-4 are within the amniotic fluid. The embryonology of all three germ layers exist in this very fluid including different subsets of specific cells that still need clarification. (A)

There are many scientists and researchers alike whose passion resides in stem cell research where they can discover new sources of stem cells. The maintenance of Oct-4 is very important to stem cells. Each mammalian pluripotent stem cell expresses within their line the Oct-4 gene, which rapidly disappears once the cells differentiate. (A)

Samples of amniotic fluid were taken from amniocentesis which is performed after the 14th week of pregnancy during routine prenatal diagnosis. Through this process, advanced maternal age and cytogenic analyses showed normal karyotypes. (A)

In routine prenatal genetic diagnosis, a wide range of fetal abnormalities which are caused by genetic alterations, are typically discovered by doctors of the patient.  Therefore, the major focus was in finding new sources of pluripotent stem cells. Here, Oct-4 acts as a marker for these very pluripotent stem cells which are expressed in embryonic carcinoma cells, embryonic stem cells, as well as embryonic germ cells. (A)

Research:

First the cells of the amniotic fluid were tested for mRNA expression of Oct-4. Along with Oct-4, stem cell factors yimentin and alkaline phosphatases through the RT+/- PCR technique were analyzed. Western blot analysis and immunocytochemistry were both used in the investigation of protein expression of Oct-4. The Oct-4-positive cells were analyzed through double immunostaining for the expression of cyclin A. (A)

Results:

The techniques that were used in this study were RT+/- PCR, Western Blot, and Immunocytochemical analyzes in order to show if in the human amniotic fluid the Oct-4- negative cells first could be found which are expresed in the nucleus. The Oct-4-positive cell samples from the amniotic fluid also expressed stem cell factors (i.e. yimentin and alkaline phosphatase mRNA). Cyclin A expression showed that these positive cells are actively dividing in the amniotic fluid.
Subsequently, these test results show that the amniotic fluid formed within the human body when a woman is pregnant can be a new source for isolating the Oct-4-positive stem cells (discussed above) avoiding ethical concerns that typically surround human embryonic research or studies. (A)

Links:
(A)http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.324.6616&rep=rep1&type=pdf

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