Mid-trimester fetal livers are a rich source of CD34+/++ cells for transplantation

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Abstract
Human fetal livers (FL), between 16 and 24 weeks of gestation, were studied for their potential as a source of hematopoietic stem cells for prenatal and postnatal transplantation. In this report we give a quantitative evaluation of human FL as a source of candidate stem cells, and develop a protocol for the isolation of these cells free of microbial contaminants and almost free of mature T cells. Human FLs contained a median 1.9 × 109 viable cells and a mean of 1.3 × 108 CD34+/++ cells (range 1.1 × 107 to 4.7 × 108). Regardless of gestational age, no significant differences were apparent in the numbers of total progenitors or in the numbers of candidate stem cells (CD34++ CD38− and CD34++CD4+), suggesting that the expansion in the liver of the early compartments of hematopoietic progenitors reaches a plateau after the sixteenth week of gestation. Colony-forming units culture (CFU-C) were found to range from 4.1 × 106 to 2.5 × 107 per FL. Positive selection of FL CD34++ cells was achieved using the Baxter Isolex 50 device. An average purity of 74% and yield of 29% of CD34+/++ cells was achieved. T cells were depleted by 99.95%, resulting in a mean of 6.5 × 103 T cells per processed liver. Analysis of candidate stem cell populations and primitive colony-forming cells (CFC) suggested a preferential enrichment of these cells over the total population of CD34+/++ cells. Processed CD34+/++cells were found to be sterile. In conclusion, purification of FL progenitors between 16 and 24 weeks of gestation results in a large number of early progenitors suitable for in utero and possibly post-natal transplantation.

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