Novel approaches to manipulating foetal cells in the maternal circulation for non-invasive prenatal diagnosis of the unborn child

Abstract

Due to the risks to the foetus with invasive prenatal diagnosis, non-invasive prenatal diagnosis (NIPD) is gaining tremendous interest but no reliable method that can be widely used has been developed to date. Manipulation of foetal cells and foetal cell-free genetic material in the maternal blood are two promising approaches being researched. The manipulation of foetal cells in the maternal circulation is more popular as it can provide complete genetic information of the foetus particularly the diagnosis of aneuploidies. However, the foetal cell numbers in the maternal circulation are small and their enrichment and ex vivo culture remain two major challenges for NIPD. Primitive foetal erythroblasts (pFEs) have been considered as a good potential candidate for early first trimester NIPD but their nature, properties and manipulation to provide adequate cell numbers remain a challenging task and several approaches need to be meticulously evaluated. In this review we describe the current status of NIPD and suggest some novel approaches in manipulating pFEs for future clinical application of NIPD. These novel approaches include (1) understanding the pFE enucleation process, (2) enriching pFE numbers by individual pick-up of pFEs from maternal blood using micromanipulation and microdroplet culture, (3) expansion of pFEs using mitogens and (4) decondensation of the pFE nucleus with histone deacetylase (HDAC) inhibitors followed by reprogramming using gene delivery protocols with/without small reprogramming molecules to improve reprogrammed pFE proliferation rates for successful NIPD.

Prenatal Diagnosis, Where and How: No Way Out?

“…Dielectrophoresis treatment does not alter the viability of manipulated cells. [44][45][46][47][48][49][50][51] …”

A Comparison of in vitro Culture of Fetal Nucleated Erythroblasts from Fetal Chorionic Villi and Maternal Peripheral Blood for Noninvasive Prenatal Diagnosis

“…The current cell culture approach used for amplifying the fetal erythroblasts from the maternal blood could not produce a sufficient number for prenatal diagnosis. Technical improvement of cell culture, such as reverting differentiated somatic cells back to the embryonic state by using nuclear reprogramming techniques [36], or the exploration of alternatives, such as extracting fetal DNA from maternal circulation [37], may be potential strategies for a breakthrough in clinical noninvasive prenatal diagnosis.…”

Intrauterine Diagnosis of Genodermatoses

“…The main challenge in this method is the ability to isolate the fetal cells from the maternal circulation, due to their very low concentrations in the maternal blood. It has been estimated that the number of fetal cells in maternal blood is 1 in 10 5 to 1 in 10 9 of mononuclear cells [45]. Additionally, the final purity of the fetal cells is low, and oftentimes they are contaminated with maternal cells [45].…”

“…It has been estimated that the number of fetal cells in maternal blood is 1 in 10 5 to 1 in 10 9 of mononuclear cells [45]. Additionally, the final purity of the fetal cells is low, and oftentimes they are contaminated with maternal cells [45]. Other obstacles include high levels of allele dropout and cellular apoptosis upon entrance to the maternal circulation [43].…”