D'Rozario James, Parisotto Robin, Stapleton Jennifer, Gidley Alison, Owen David
Bone Marrow Transplant Laboratory, ACT Pathology, Garran, ACT 2605, Australia; Australian National University Medical School, Canberra, ACT 0200, Australia; Bone Marrow Transplant Unit, Clinical Haematology Department, Canberra Hospital and Health Services, Garran, ACT 2605, Australia.
Bone Marrow Transplant Laboratory, ACT Pathology, Garran, ACT 2605, Australia.
Transfus Apher Sci. 2014 Jun;50(3):443-50. doi: 10.1016/j.transci.2014.02.021. Epub 2014 Mar 12.
By convention, peripheral blood stem cell products for autologous transplantation are evaluated for quality by CD34(+) cell dose at the time of harvesting. A CD34(+) cell dose in excess of 2.0 × 10(6)/kg of recipient body weight is considered adequate for haematopoietic engraftment. Viable CD34(+) cell numbers are enumerated in most laboratories using the ISHAGE single platform flow cytometric method which utilizes monoclonal antibodies to CD45, CD34 and 7 amino actinomycin D (7AAD) dye exclusion.
One hundred and six consecutive autologous transplantation procedures underwent viable CD34(+) cell enumeration at the time of harvesting and post thaw prior to re-infusion. Neutrophil and platelet engraftment and markers of haematopoietic support were analyzed.
Mean pre-cryopreservation viable CD34(+) numbers were 4.882 × 10(6)/kg. Mean post thaw viable CD34(+) numbers were 3.234 × 10(6)/kg. Mean loss of viable CD34(+) cells with processing and cryo-preservation was 1.648 × 10(6)/kg (33%). For neutrophil engraftment, there was no significant difference between high (⩾ 3.0 × 10(6)/kg) and low (<1.5 × 10(6)/kg) post thaw viable CD34(+) cell counts (p=0.545). For platelet engraftment, there was however a significant difference observed between the high and low pre infusion viable CD34(+) groups (p<0.001). Additionally, significant differences were seen between the post thaw viable CD34(+) cell count and the associated length of hospital admission, days of use of G-CSF post transplantation, use of antibiotics in the post transplantation period and transfusion support in the post transplantation period.
A significant loss of viable CD34(+) cells occurs during processing, cryopreservation and thawing. Low numbers of viable CD34(+) cells infused post thaw will still result in adequate neutrophil engraftment however may delay platelet engraftment. Low viable CD34(+) cell numbers have significant effects on admission duration and use of haematopoietic supportive measures with consequent effects on healthcare resources.
按照惯例，自体移植的外周血干细胞产品在采集时通过CD34(+)细胞剂量来评估质量。接受者体重每千克超过2.0×10(6)个CD34(+)细胞的剂量被认为足以实现造血植入。大多数实验室使用ISHAGE单平台流式细胞术方法来计数活的CD34(+)细胞数量，该方法利用针对CD45、CD34的单克隆抗体以及7氨基放线菌素D（7AAD）染料排除法。
对106例连续的自体移植手术在采集时以及解冻后再输注前进行活的CD34(+)细胞计数。分析中性粒细胞和血小板植入情况以及造血支持标志物。
冻存前活的CD34(+)细胞数量均值为4.882×10(6)/kg。解冻后活的CD34(+)细胞数量均值为3.234×10(6)/kg。处理和冻存过程中活的CD34(+)细胞的平均损失为1.648×10(6)/kg（33%）。对于中性粒细胞植入，解冻后活的CD34(+)细胞计数高（⩾3.0×10(6)/kg）和低（<1.5×10(6)/kg）组之间无显著差异（p = 0.545）。然而，对于血小板植入，输注前活的CD34(+)细胞高、低组之间观察到显著差异（p<0.001）。此外，解冻后活的CD34(+)细胞计数与相关的住院时间、移植后使用G-CSF的天数、移植后使用抗生素情况以及移植后输血支持之间存在显著差异。
在处理、冻存和解冻过程中活的CD34(+)细胞发生显著损失。解冻后输注的活的CD34(+)细胞数量低仍将导致足够的中性粒细胞植入，但可能会延迟血小板植入。活的CD34(+)细胞数量低对住院时间和造血支持措施的使用有显著影响，从而对医疗资源产生影响。
