A modified harvest technique for cord blood hematopoietic stem cells.

Human umbilical cord blood is an excellent source of hematopoietic stem cells for research and bone marrow transplantation. We have developed a modified technique which effectively flushes and collects placental derived stem cells. Catheters were inserted into the umbilical artery and vein. Using this sterile and closed system technique, 10 saline diluted cord blood harvests had hematocrits of 16% to 25%, total mononuclear cell counts of 1.17 to 34.4 x 10(8) and volumes of 105-245 ml. The averaged extrapolated colony forming units from five cord blood harvests were as follows: CFU-GM 5.2 x 10(5) (9.96 x 10(4) to 1.24 x 10(6)), BFU-E 8.82 x 10(5) (2.40 x 10(4) to 1.61 x 10(6)) and CFU-GEMM 1.37 x 10(5) (7.33 x 10(3) to 3.18 x 10(5)). This quick technique avoids needle exposure and collects a significant volume of stem cells.

Midazolam versus fentanyl as premedication for painful procedures in children with cancer.

Premedication for painful procedures in children with cancer is not routinely used. Many medications used are only intermittently effective or require special equipment or anesthesia support. In a randomized, double-blind, crossover study, the safety and efficacy of midazolam, a short-acting benzodiazepine, were compared with the safety and efficacy of fentanyl, a short-acting narcotic analgesic. In 25 children studied, 100% of children and their parents preferred study drugs to any previous premedication. Seventy-two percent preferred midazolam to fentanyl. Preprocedural anxiety, adverse behavioral symptoms, and visual analog scales all improved and side effects were minimal. It is concluded that premedication for painful procedures should be used routinely in children with cancer. With proper monitoring, fentanyl and midazolam can be used safely in the outpatient clinic setting. Midazolam was found to be the drug of preference for the majority of patients.

Myeloablative chemoradiotherapy and autologous bone marrow infusions for treatment of neuroblastoma: factors influencing engraftment.

Bone marrow harvested from cancer patients for autologous bone marrow reinfusion (ABMR) after myeloablative treatment may be injured, in both its proliferating and stromal cell pools, by either previous treatment or manipulation at the time of harvest. We have examined the relative effects of seven covariates on hematologic recovery after ABMR in children with neuroblastoma (NBL) using univariate and step-up analysis. We measured recovery by times to achieve (1) white blood cell counts greater than 1,000/microL; (2) absolute neutrophil counts greater than 500/microL; and (3) platelet counts greater than 20,000/microL without transfusion. In univariate analysis, recovery was significantly associated with the amount of prior chemotherapy and the interval between last chemotherapy and marrow harvest. Patient sex, the number of granulocyte-macrophage colonies infused, harvest-to-freeze interval, and use of purging were marginally associated. After adjusting for potential confounders in a multivariate model, the amounts of chemotherapy and granulocyte-macrophage colonies infused were independently significant predictors of time to total white blood cell count recovery; chemotherapy courses and chemo-to-harvest interval were predictors of neutrophil count recovery; and sex, use of purging, and harvest-to-freeze interval were marginal predictors of platelet recovery. The speed of hematologic recovery after ABMR seems to depend mainly on pre-existing factors and marginally on manipulation of the marrow after harvest. These factors may affect both proliferating and stromal cell pools.

Transporting bone marrow for in vitro purging before autologous reinfusion.

We describe the procedures employed for transporting bone marrow to and from a central facility. Marrow has been harvested from 80 patients with neuroblastoma, at 16 centers which are geographically dispersed throughout North America. Marrow from the outside transplant centers was packed on wet ice or cold packs in insulated containers, and transported by commercial carriers or chartered aircraft to the central processing laboratory. Post processed marrows were frozen in liquid nitrogen and returned by commercial carrier to the referring institution. In comparing transported with non-transported but similarly treated marrows, no differences were found in any of the following parameters: (1) CFU-GM recovery, (2) fraction viable cells at thawing, or (3) time to engraftment in patients. We conclude the transportation of harvested marrows to a central purging facility is safe. Based on this experience, we propose a set of standards, which, if adhered to, will insure the continued safe processing, shipping, and storage of bone marrow in all centers so engaged.