Human CD34+ cell isolation from fetal liver, and fetal thymus preparation v1

This protocol details the steps for isolating human CD34+ cells from human fetal liver. It also explains how to prepare human fetal thymus for immediate use or for freezing, as well as the process for thawing. The CD34+ cells are hematopoietic progenitor cells and can be used to generate humanized mice through reconstitution of immune cells via IV injection after bone marrow ablation. These cells can also be used for mixed lymphocyte reaction experiments.

CD34+ isolation from human bone marrow v1

This protocol details the steps for isolating CD34+ cells from human bone marrow. The CD34+ cells isolated from this protocol can be used for generating humanized mice through reconstitution of immune cells via IV injection after bone marrow ablation. These cells can also be used for mixed lymphocyte reaction experiments.

Intramedullary injury combined with osteoporosis therapeutics regulates targeted local osteogenesis

Bone marrow ablation prompts transient bone formation in nearly the entire medullary cavity before marrow regeneration occurs. Here, we establish a procedure to direct bone formation in a desired particular site within the medullary cavity for support of biomedical devices. Local intramedullary injury was performed in the tibiae of rats and parathyroid hormone (PTH), alendronate, or saline was administered. Newly generated bone in the medulla was assessed by micro-CT and histology. To evaluate the function of newly generated bone, animals received intramedullary injury in tibiae followed by daily PTH. At day-14, implants were placed in the endocortical bone and the bone response to the implants was assessed. The fate of newly generated bone was compared with and without implants. We found that neither intramedullary injury nor medication alone resulted in bone formation. However, when combined, substantial bone was generated locally inside the diaphyseal medulla. Newly formed bone disappeared without implant placement but was retained with implants. Bone was especially retained around and between the implants. This study found that local bone marrow disruption followed by PTH or alendronate generated substantial cancellous bone locally in the diaphyseal medulla. This approach offers promise as a tissue engineering tool in medicine and dentistry.

Preliminary dosimetric evaluation of 90Y-BPAMD as a potential agent for bone marrow ablative therapy

AimBone-seeking radiopharmaceuticals are potential therapeutic tools for bone marrow ablation in patients with multiple myeloma. In this procedure, estimation of radiation absorbed dose received by the target and non-target organs is one of the most important parameters that should be undertaken. This research revolves around the absorbed dose to human organs after 90Y-BPAMD injection.Materials and methods90Y-(4-{[(bis(phosphonomethyl))carbamoyl]methyl}-7,10-bis(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl) acetic acid (90Y-BPAMD) complex was successfully prepared under optimised conditions. The human absorbed dose of the complex was estimated based on the biodistribution data on rats using the radiation-absorbed dose-assessment resource method. The target to non-target absorbed dose ratios for the complex was compared with the ratios for 166Ho-DOTMP, as the main radiopharmaceutical for bone marrow ablation.ResultsAs expected, the highest amounts of absorbed dose were observed in the bone surface and the bone marrow with 2·52 and 2·29 mGy/MBq, respectively. The red marrow to the most organ absorbed dose ratios for 90Y-BPAMD are much higher than the ratios for 166Ho-DOTMP.Findings90Y-BPAMD has interesting characteristics compared with 166Ho-DOTMP and can be considered as a high potential agent for bone marrow ablative therapy of the patient with multiple myeloma.