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Much of our current knowledge on the in vitro proliferation and differentiation of HPCs—particularly those from BM—has derived from studies using Dexter-type long-term cultures (D-LTC) . In this experimental system hematopoietic cell growth occurs in close association with an adherent cell layer formed by marrow stromal elements, which are derived from the same marrow sample. Under these culture conditions, hematopoiesis can be sustained for several weeks, or even months, without the addition of exogenous cytokines, since marrow stromal cells produce both extracellular matrix proteins and a variety of cytokines necessary for the development of hematopoietic cells [18-20].
We have previously shown that when human umbilical cord blood (UCB) cells are cultured in standard Dexter-type long-term cultures (D-LTC), adherent cells develop forming a discrete net on the bottom of the culture flask. The identity of such cells, however, has not been defined. Accordingly, the major goal of the present study was to characterize the adherent cells developed in standard UCB D-LTC. Cultures were established from 14 UCB samples and from nine bone marrow (BM) samples, as controls.
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Kinetics of Hematopoiesis in Dexter-Type Long-Term CulturesEstablished from Human Umbilical Cord Blood CellsHECTOR MAYANI,aMARGARITA GUTIÉRREZ-RODRÍGUEZ,aLAURA ESPINOZA,aEDITH LÓPEZ-CHALINI,bALEJANDRA HUERTA-ZEPEDA,aEUGENIA FLORES,aELIZABETH SÁNCHEZ-VALLE,aFERNANDO LUNA-BAUTISTA,aIGNACIO VALENCIA,aOCTAVIO T. RAMÍREZbaOncological Research Unit, Oncology Hospital, National Medical Center, IMSS, Mexico City, Mexico;bBioengineering Department, Institute of Biotechnology, UNAM, Cuernavaca, MexicoKey Words.
To determine whether these processes are dependent upon the same or different populations of stromal cells, stromal cell lines were generated from the adherent layer of a Dexter type long-term bone marrow culture. These cultures support myeloid cells and their precursors, a B cell precursor, and the adherent layer cells with support B cell differentiation under appropriate conditions.