Serum supplement, inoculum cell density, and accessory cell effects are dependent on the cytokine combination selected to expand human HPCs ex vivo.

BACKGROUND: The prolonged periods of pancytopenia associated with cord blood transplants suggest that in some cases cell numbers may be limiting. The possibility that limiting cell numbers may be overcome and prolonged periods of pancytopenia abrogated by the transplantation of human umbilical cord blood cells expanded ex vivo has led to efforts to define optimal culture conditions for these cells. STUDY DESIGN AND METHODS: Cord blood CD34+ cells were cultured with three cytokine combinations: SCF+G-CSF+GM-CSF+MGDF (SGGM); IL-6+ SCF+MGDF+Flt3-ligand (6SMF); and IL-1+IL-3+IL-6+G-CSF+GM-CSF+SCF+Epo (GFmix). Serum effects, inoculum concentration (cells/mL) seeding density (cell/cm(2)) and accessory cell effects on the expansion of CD34+ cells were determined. RESULTS: Cellular outputs were significantly higher with fetal calf serum (FCS) than with cord blood serum (CBS) or adult group AB serum (ABS) in the presence of 6SMF, however, CBS was as effective as FCS. The best seeding concentrations varied for each of the cytokine combinations, and inoculum densities exceeding 1000 cells per cm(2) proved detrimental for cultures containing GFmix and SGGM. Accessory cell studies indicated that populations expressing the CD33 antigen inhibited the expansion of purified CD34+ cells in the presence of GFmix or SGGM, but not in the presence of 6SMF. CONCLUSION: Serum supplement, inoculum cell concentration, seeding densities, and accessory cell effects are dependent upon the cytokine combination selected to expand cord blood HPCs ex vivo. Thus, each of these measures should be assessed to establish reproducible and reliable conditions for the selection of different cytokine combinations to culture cord blood HPCs.

Cytogenetic studies in endometriosis tissue.

We analyzed 45 pelvic endometrial implants from 42 patients between the ages of 14 and 40 years to investigate whether cytogenetic abnormalities were present in these samples. Chromosomal abnormalities have been described in benign tumors of the female genital tract, such as uterine leiomyomas and endometrial polyps. Furthermore, heritable factors have been postulated to influence a woman's susceptibility to develop endometrial tissue implants outside the uterine cavity. To study whether these factors result in or are associated with chromosomal changes, endometriosis cells were analyzed cytogenetically after short-term culture. No consistent chromosome abnormalities were observed in any of the cases analyzed.

Nonrandom cytogenetic changes in leiomyomas of the female genitourinary tract. A report of 35 cases.

Cytogenetic analysis of short-term cultures from 35 leiomyomas of the female genitourinary tract showed abnormal karyotypes in 14 cases. In 11 of 14 aberrant tumors, normal cells were also observed. Structural changes were most frequent, resulting in modal chromosome numbers in the diploid range. Our data confirm preferential breakpoint clusters at 7q, 12q14-15, and 14q23-24, mainly resulting from consistent, specific chromosome rearrangements such as t(12;14)(q14-15;q23-24) and del(7)(q21) or del(7)(q22q32). Together with previously published cases, we describe trisomy 12, ring chromosomes, and monosomy 22 as new additional recurrent findings in myomas. Statistical analyses of possible coherencies between tumor karyotype (abnormal versus normal) and clinicopathologic data, as well as age of the patients, menopausal status, and tumor size showed no correlations.

Cytogenetic findings in nine leiomyomas of the uterus.

Chromosomal analysis of nine benign leiomyomas of the uterus after short-term culture showed karyotypic abnormalities in four cases. All four exhibited multiple chromosome changes, including three cases characterized by complex chromosome rearrangements involving a number of chromosomes. Among others, these rearrangements included a translocation between chromosomes 12 and 14 in one case, a deletion of chromosome 7q in two cases, and both del(7q) and a complex translocation involving chromosomes 12 and 14 in another case. These results confirm the involvement of chromosomes 7, 12, and 14 in leiomyomas and indicate that benign tumors can also be characterized by complex cytogenetic changes.

inv(12)(p11.2q13) in an endometrial polyp.

A benign endometrial polyp from a 50-year-old postmenopausal woman has been cytogenetically investigated. A single clonal karyotypic anomaly, inv(12)(p11.2q13), was found in about 30% of cells analyzed after short-term culture. This finding contributes further to the hypothesis that the chromosomal segment 12q13-q14, which is also involved in chromosomal rearrangements in uterine leiomyomas, pleomorphic adenomas of the salivary glands, lipomas, and myxoid liposarcomas, contains a gene or genes that are related to cellular proliferation rather than to malignant transformation.