Criswell Kay A, Bock Jeffrey H, Johnson Kjell, Criswell Ryan A, Giovanelli Richard P
Westbrook Biomarker and Pharma Consulting, Westbrook, Connecticut.
Pfizer Worldwide Research and Development, Drug Safety Research and Development, Groton, Connecticut.
Vet Clin Pathol. 2018 Dec;47(4):539-555. doi: 10.1111/vcp.12672. Epub 2018 Nov 26.
In a previous study, the validation of rat bone marrow (BM) collection, processing, and analysis using the Sysmex XT-2000iV (Sysmex Corporation, Kobe, Japan) hematology analyzer showed that the Sysmex hematology analyzer produced BM differential counts that were comparable to those obtained with microscopic differential counts.
This study was conducted to expand the validation of the Sysmex TNCC (total nucleated cell count) and 5-part BM differential in cynomolgus monkeys, Beagle dogs, and CD-1 mice, which are alternate species that are also frequently used in preclinical safety studies.
The Sysmex 5-part BM differential counts were generated with a two-step process, whereby proliferating and maturing erythroid and myeloid cells were determined by preset gating and lymphocytes were determined using species-specific B- and T-lymphocyte antibodies and a magnetic cell-sorting method (MACS). Agreement with microscopic myelograms with 500-cell differential counts was determined from BM suspensions of 62 cynomolgus monkeys, 47 Beagle dogs, and 44 CD-1 mice.
The correlation coefficients between methods for myeloid to erythroid (M:E) ratios in all three species was > 0.928. The Bland-Altman differences between methods were approximately ± 0.3 units for the M:E ratio in dogs and mice, and +0.6 and -0.4 in monkeys. The upper limits of agreement for all three species were ≤7% for maturing myeloid cells, ≤6% for maturing erythroid cells, and ≤4% for proliferating myeloid cells, proliferating erythroid cells, and lymphocytes.
The Sysmex XT-2000iV produces an automated M:E ratio and a 5-part differential count equivalent to microscopic differential counts in cynomolgus monkeys, Beagle dogs, and CD-1 mice.
在之前的一项研究中，使用Sysmex XT - 2000iV（Sysmex公司，日本神户）血液分析仪对大鼠骨髓（BM）的采集、处理及分析进行验证，结果表明Sysmex血液分析仪得出的骨髓分类计数与显微镜下分类计数结果相当。
本研究旨在扩大对食蟹猴、比格犬和CD - 1小鼠中Sysmex总核细胞计数（TNCC）和五分类骨髓分类的验证，这些替代物种也常用于临床前安全性研究。
Sysmex五分类骨髓分类计数通过两步法得出，即通过预设门控确定增殖和成熟的红系及髓系细胞，并使用物种特异性B和T淋巴细胞抗体及磁珠细胞分选法（MACS）确定淋巴细胞。从62只食蟹猴、47只比格犬和44只CD - 1小鼠的骨髓悬液中确定与500细胞分类计数的显微镜骨髓象的一致性。
所有三个物种中髓系与红系（M:E）比值的方法间相关系数均>0.928。犬和小鼠中M:E比值的方法间布兰德 - 奥特曼差异约为±0.3个单位，猴中为 +0.6和 -0.4。所有三个物种中成熟髓系细胞的一致性上限≤7%，成熟红系细胞≤6%，增殖髓系细胞、增殖红系细胞和淋巴细胞≤4%。
Sysmex XT - 2000iV在食蟹猴、比格犬和CD - 1小鼠中产生的自动化M:E比值和五分类计数等同于显微镜下分类计数。
