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Derivation of Human Embryonic Germ Cells: An Alternative Source of Pluripotent Stem Cells

^a Division of Human Genetics and
^b Maternal, Fetal, and Neonatal Physiology Group, Fetal Origins of Adult Disease Division, University of Southampton, Southampton General Hospital, Southampton, United Kingdom

Key Words. Pluripotent • Embryonic stem • Primordial germ cells • Embryonic germ • Embryoid body • Differentiation

Neil A. Hanley, MB.Ch.B., Ph.D. and Lee Turnpenny, Ph.D., Division of Human Genetics, University of Southampton, Duthie Building (MP 808), Southampton General Hospital, Tremona Road, Southampton SO16 6YD, United Kingdom. Telephone: 44-0-23-8079-6421; Fax: 44-0-23-8079-4264; e-mail: N.A.Hanley{at}soton.ac.uk, lturnpen{at}hgmp.mrc.ac.uk

Based on evidence suggesting similarities to human embryonic stem cells, human embryonic germ (hEG) cells have been advocated as an alternative pluripotent stem cell resource but have so far received limited attention. To redress this imbalance, human fetal gonads were collected for the isolation and culture of primordial germ cells at 7-9 weeks postconception. We provide evidence for the derivation, culture, and differentiation of hEG cells in vitro. This evidence includes the expression of markers characteristic of pluripotent cells, the retention of normal XX or XY karyotypes, and the demonstration of pluripotency, as suggested by the expression of markers indicative of differentiation along the three germ lineages (ectoderm, mesoderm, and endoderm) and an associated loss of pluripotent markers. In assessing this differentiation, however, we also demonstrate a hitherto unacknowledged overlap in gene expression profiles between undifferentiated and differentiated cell types, highlighting the difficulty in ascribing cell lineage by gene expression analyses. Furthermore, we draw attention to the problems inherent in the management of these cells in prolonged culture, chiefly the difficulty in preventing spontaneous differentiation, which hinders the isolation of pure, undifferentiated clonal lines. While these data advocate the pursuit of pluripotent hEG cell studies with relevance to early human embryonic development, culture limitations carry implications for their potential applicability to ambitious cell replacement therapies.

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