Density gradient separation of peripheral blood stem cells: comparison of an automated cell processing device and manual methods

Peripheral blood stem cells were collected from normal donors by leukapheresis on a cell separator. The leukapheresis product contained 1.5 x 10(10) mononuclear cells (MNCs) and was divided into two aliquots that underwent either automated or manual density gradient separation with ficoll-hypaque and subsequent washing. In the automated process, recovery of MNCs was 85 percent, reduction in platelet content was 64 percent, and the final hematocrit (Hct) was less than 1 percent. The manual separation resulted in 76-percent MNC recovery, a 79-percent reduction in platelet content, and a final Hct of less than 1 percent. The purified MNCs were then placed in methylcellulose culture at a concentration of 4 x 10(5) MNCs per mL. Quadruplicate 1-mL aliquots were cultured, and colonies were counted and classified on Day 14. Comparison of automated and manual ficoll-hypaque separations demonstrated no differences in the total, erythroid, or granulocyte-macrophage colony numbers. The cell processor used is fast, reliable, uncomplicated, and provides a sterile product containing progenitor cells that are not adversely affected by the automated ficoll-hypaque separation.     

A functional comparison of CD34 + CD38- cells in cord blood and bone marrow

We present cell cycling and functional evidence that the CD34+CD38- immunophenotype can be used to define a rare and primitive subpopulation of progenitor cells in umbilical cord blood. CD34+CD38- cells comprise 0.05% +/- 0.08% of the mononuclear cells present in cord blood. Cell cycle analysis with the fluorescent DNA stain 7- aminoactinomycin D showed that the percentage of CD34+ cells in cycle directly correlated with increasing CD38 expression. CD34+CD38- cord blood cells were enriched for long-term culture-initiating cells (LTCIC; cells able to generate colony-forming unit-cells [CFU-C] after 35 to 60 days of coculture with bone marrow stroma) relative to CD34+CD38- cells. In an extended LTCIC assay, CD34+CD38- cells were able to generate CFU-C between days 60 and 100, clearly distinguishing them from CD34+CD38+ cells that did not generate CFU-C beyond day 40. When plated as single cells, onset of clonal proliferation was markedly delayed in a subpopulation of CD34+CD38- cells; clones (defined as > 100 cells) appeared after 60 days of culture in 2.9% of CD34+CD38- cells. In contrast, 100% of CD34+CD38+ cells formed clones by day 21. Although the CD34+CD38- immunophenotype defines highly primitive populations in both bone marrow and cord blood, important functional differences exist between the two sources. CD34+CD38- cord blood cells have a higher cloning efficiency, proliferate more rapidly in response to cytokine stimulation, and generate approximately sevenfold more progeny than do their counterparts in bone marrow.     

An interactive CD-ROM to inform patients about stem cell transplantation

OBJECTIVE: Cancer patients receiving chemotherapy or a Stem Cell Transplantation (SCT) are in need of information about their disease, treatment options and side effects. Patient education usually has to be given within limited time. Under these circumstances, patients may find it difficult to completely understand and to retain the information given. METHODS: As a supplement to standard information methods we developed an interactive CD-ROM with information on SCT. This CD-ROM provides both medical information and more subjective patients' experiences. Part one provides information regarding the treatment course from diagnosis through to post-discharge care. The second part consists of interviews with former patients and describes their experiences. As the system is interactive, it can be utilised according to the patient's individual preferences. The CD-ROM comprises audio, video, animations, pictures, and text. Printing of certain sections is optional. The technical format of the CD-ROM makes it relatively simple to utilise the information and to make it suitable for other institutions or even other treatments. In this preliminary study the acceptability of the interactive CD-ROM by patients undergoing a SCT is described. RESULTS: Patients' overall evaluations of the interactive CD-ROM were highly positive. For example, 90.2% (N=51) found it interesting, clear, useful and valued getting information by means of a CD-ROM. Most patients would recommend the interactive CD-ROM to other patients in the same situation. CONCLUSION: The content of the CD-ROM on SCT as well as the computer-based interactive method are well accepted by patients. PRACTICE IMPLICATIONS: Computer-based education may enhance patient education and thus the quality of patient care. We must now establish the program's effectiveness. Moreover, plans have been developed to disseminate the information on SCT over the Internet. Future development of comparable programs and their evaluation should be encouraged to promote the well-being of cancer patients.