Evaluation of the embryonic preimplantation potential of human adult somatic cells via an embryo interspecies bioassay using bovine oocytes

Objective

To examine the embryonic preimplantation potential of human adult somatic cells by creating interspecies embryos via somatic cell nuclear transfer (SCNT) using bovine oocytes.

Design

Prospective study.

Setting

Research facility of Reprogen.

Patient(s)

Infertile couples.

Intervention(s)

Enucleated bovine oocytes were fused via SCNT with either human granulosa (HG) or fibroblast (HF) cells and cultured in vitro. Polymerase chain reaction (PCR) and DNA analysis were performed on the interspecies embryos. Parthenogenetically activated embryos served as controls.

Main Outcome Measure(s)

Embryonic preimplantation development after interspecies SCNT.

Result(s)

From enucleated bovine oocytes fused with HG cells (n = 48) and HF cells (n = 75), 15 HG- and 22 HF-derived embryos developed, some of which progressed to blastocysts (31.3% vs. 29.3%, respectively). The PCR and DNA analysis showed that the interspecies embryos contained human genomic DNA specific for the individual DNA profile of the HG or HF donor cells used for SCNT. In addition, both bovine- and human-specific mitochondrial DNA was detectable in the interspecies embryos up to the blastocyst stage. Parthenogenetic development was 46.8% and 64.9% for the HG and HF series, respectively. The SCNT efficiency index, defined as the ratio of SCNT and parthenogenetic success rate, was 66.8% for HG cells and 45.5% for HF cells.

Conclusion(s)

This interspecies bioassay can be utilized to determine and assess the embryonic preimplantation potential of different types of human adult somatic cells.

Key Words:  Human granulosa cell , human fibroblast , bovine oocyte , interspecies SCNT , microPCR , DNA sequencing