[Abstract]  [Full Text]  [PDF]

Marcus O. Muench^1*, David L. Suskind¹ and Alicia Bárcena¹

¹ Department of Laboratory Medicine, University of California at San Francisco. 3rd & Parnassus Ave., Room U-440; San Francisco, CA 94143-0793. . Phone: (415) 476-8420 Fax: (415) 476-2956

* To whom correspondence should be addressed: Marcus Muench, Department of Laboratory Medicine, University of California at San Francisco. 3rd & Parnassus Ave., Room U-440; San Francisco, CA 94143-0793. Phone: (415) 476-8420. Fax: (415) 476-2956. Email: muench@itsa.ucsf.edu

Biol. Proced. Online 2002;4:10-23. doi:10.1251/bpo29
Submitted: April 16, 2002; Accepted: May 27, 2002; Published: June 11, 2002.

Indexing terms: fetal tissue; hematopoietic stem cells; cell differentiation; natural killer cells; dendritic cells.

The study of hematopoietic stem cells (HSCs) and the process by which they differentiate into committed progenitors has been hampered by the lack of in vitro clonal assays that can support erythroid, myeloid and lymphoid differentiation. We describe a method for the isolation from human fetal liver of highly purified candidate HSCs and progenitors based on the phenotypes CD38^-CD34⁺⁺ and CD38⁺CD34⁺⁺, respectively. We also describe a method for the growth of colony-forming cells (CFCs) from these cell populations, under defined culture conditions, that supports the differentiation of erythroid, CD14/CD15⁺ myeloid, CD1a⁺ dendritic cell and CD56⁺ NK cell lineages. Flow cytometric analyses of individual colonies demonstrate that CFCs with erythroid, myeloid and lymphoid potential are distributed among both the CD38^- and CD38⁺ populations of CD34⁺⁺ progenitors.