フローサイトメトリーによるCD34陽性細胞 検出に関するガイドライン
Contents
1. 1 2 025 CD34 S8C CD34 CD34 CD342. 5 5 3 5 5 2 1 11 5 5 4 CD34 5 5 1 5 5 3 CD34 CD34 NN NI CD34
3. 0 5 2 PBS 1 100 ug L 4 3 4 3 1 3 1 18 22 15 30
4. 3 2 CD34 CD34 CD34 Pasteurella haemolytica 0 CD34 3 5 1 CD34 My10 Immu 133 Immu 409 BI 3C5 I FITC
5. 5 CD34 CD34 ISCT ISHAGE 3 CD34 6 9 10 ISHAGE 5 1 FSC SSC CD45 CD34 4 2 5 3
6. 6 5 6 3 1 2 CD34 0 01 CD34 CD45 1 75000 2 15 CD34 100 3 6 5 6 4
7. 0 HUK 488nm CN 068 2 CD45 CD34 5 4 CD34 5 4 1 FSC 8SC CD45 CD34
8. 6 6 1 CD34bight CD34dim CD34bright CD34dim OD45 FSC SSC 6 2 CD34
9. 3 OD34 PT 72 D 7 AAD TTITC CD45 PE CD34 PE FL2 7 AAD JOCLS H2 P DNA ProCOUNT viability
10. 5 3 2 CD34 CD34 OCD34 4 1 JCCLS H1 A JCCLS H2 P 9 viability se Cc CD34 3 4 4 2 WBC 1 2000 20000
11. 00 5 3 7 PBS Ca Mg Hank s BS8 0 5 2 CD34 5 3 2 5 4 CD34
12. JCCLS Japanese Commnittee for Chnical Laboratory Standards 2 JCCLS H1 A JCCLS H2 P approve public comment approve CD34
13. 2 1 2 2 1 3 2 2 2 4 22 JCCLS H1 A 2 3 37 40 2 3 2 3 1
14. 40 gm 3 CD34 CD34 CD45 2 3 4 ISHAGE International Society for Hematotherapy and Graft Engineering 3 3 1 4 3 3
15. CD34 CD45 OD34 CD45 CD34 CD45dim CD34 CD45bright CD34 CD457 SSC CD45 CD34 DNA ProCOUNTTM CD45 CD45 3 CD34 3 4 3 4
16. 1
17. CD34 CD34 CD34 2 2 1 2 1 1 G CSF PBSCH Peripheral Blood Stem Cell Harvest
18. 2 8 PI 4 3 2 CD34 CD34
19. 5 12 CD34 CD34 ww 7 1 161 410 412 1992 2 151 65 66 1989 3 Sutherland DR Anderson L Keeney M et al The ISHAGE guidelines for CD34 cells determination by flow cytometry J Hematotherapy 5 213 226 1996 4 Schlossman SF et al Ed Leukocyte typing V Oxford University Press London 1995 5 Kishimoto T et al Ed Leukocyte typing VI Garland Publishing In
20. QBEnd 10 ICH3 NU 4A1 CLB MD34 2 8G12 581 563 BIRMA K3 CD34 1 PE I 488nm PE 3 3 CD45 CD45 OCD45 CD34 2 CD34 OD45
21. 5 6 4 A B CD34 5 2 JCCLS H2 P 5 3 CD34 5 93 1 CD34 CD45 CD34 CD34 WBC
22. CD34 CD34 CD34 3 5 CD34 CD34 8 CD34 CD34 CD34 CD45
23. 6 1 CD34bight CD34dim 6 2 6 3 CD34 6 4 6 5 7 8 20
24. CD34 ux L CD34 x WBC 7 pL CD34 7 CD45 Xx WBC 7g L Xx CD34 CD45 5 3 2 CD34 7g LD CD34 x 7 x L CD45 CD34
25. CD34 JCCLS H3 P V1 0 KReport of the Hematology Standardization Committee uidelines for CD34 Cell Determination by Flow Cytometry JCCLS H3 P V1 0 Japanese Committee for Clinical Laboratory Standards JCCLS Area Committee on Hematology Subcommittee on Flow Cytometry JCCLS Proposed Guideline JCCLS REAX jccls jccls org FAX 03 3669 9112 3 2 CD34 3 3 CD45 3 4 3 5 1 3 6 2 3 7 2 1 4 2 1 1 4 1 4 2 2 1 2 4 3
26. logical gating CD34 5 4 1 a d SSC CD45 CD45 5 4 1 a SSC CD34 CD34 5 4 1 b CD34 SSC CD45 5 4 1 d SSC CD45dim SSC FSC 5 4 1 c SSC ESC CD34 CD45dim SSC FSC C d CD45 AND CD34 D45 AND CD34 AND CD45dim CD45Sdim CD45S FITC CD34 PE HPC 3 361 34 HPC 536 uL 1 1 1 9 8 1 18 1 B 1 BB 18 1 B 1000 a 200 a00 6 gaa1iaag CD45 FITC CD34 PE CD45 FITC FS Side Scatter gn dh GoD son 1nm ide Scatter gn dn GoD Son 1nm Side Scatter gn dn GOD Son 1nm Side Scatter gn dh Gon Bm 1 O TI O O w w 10 1n 1 1 n 1n 1n 1n 1n 1n 1 el dn Gon BS Ton Chd5 FITC Cd FE Chrd5 FITC Fnmward Soatter 5 4 1
27. CD34 CD45dim 3 6 CD34 1 PBSCH Peripheral Blood Stem Cell Harvest 0 2 PBSCH 3 CD45 Se 4 CD34
28. 2 1 3 4 3 1 2 2 4 3 2 2 3 4s0 9 2 3 1 4 3 4 3 5 3 1 5 1 5 2 5 3 CD34 5a3r1 82 0 93 8 5 4 5 4 1 5 4 2 Se4 39 6 5 5 5 5 1 Threshold Noise Discrimination 5 5 2 5 5 3 5 5 4 CD34 5 6 5 6 1 5 6 2 5 6 3 5 6 4 95 6 5 6
29. PBSCH BONE MARROW CORD BLOOD a SSC CD45 SSC b SSC CD34 CD45 c SSC CD45 CD45 CD34 d FSC SSC CD34 CD45dim 5 4 2 5 2 ET EN PI 7 AAD 6 SSC CD34 5 4 3 CD34 1 Sd FE 5 4 2 SSC 7 AAD CD34 7 AAD Ungated TIME Oh mtnF Oh mnFltnF
30. 5 6 4 6 3 CD34 OCD34 5 5 4 CD34 6 4 CD34 L CD34 6 5
31. CD34 5 6 5 6 1 RT 1 ia a 5 6 2 CD34 CD45 2 CD34 CD45 CD34 CD45 CD34 CD34
32. Gate TruCOUNT beads nhI 8 12 15 gh ht 8 12 th gh gi dnn ill 1 dun kill 1 Time 12 00 ga Time m12 00 ga 5 1 CD45 0 5 4 3 5 5 10 FSC SSC Threshold Noise Discrimination MM 5 5 1 Threshold Noise Discrimi nation CD34 FSC
33. R1 7074 CD34 CD34 cells R6 493 cells gw L CD34 cells L CD34 493 7074 x 1000 gL Xx 1 70 TruCOUN 50000 50 p LL 1000 pL
34. New York amp London 1997 6 Barnett D Janossy G Lubenko A et al Guideline for the flowcytometric enumera tion of CD34 haematopoietic stem cells Prepared bythe CD34 haematopoietic stem cell working party General Haematology Task Force of the British Committee for 13 Standards in Haematology Clin Lab Hae matology 21 301 308 1999 7 JCCLS JCCLS H1 A V2 0 15 1243 136 2000 8 JCCLS PU oe JCCLS H2 P V1 0 18 69 107 2003 9 Gratama JW Orfao A Barnett D et al Flow cytometric enumeration of CD34 hematopoietic stem and progenntor cells European Working Group on Clinical Cell Analysis Cytometry 34 128 142 1998 10 Brocklebank AM and Sparrow RL Enu meration of CD34 cells in cord blood a variation on a single platform flow cyto metric method based on the ISHAGE gating strategy Cytometry 46 254 261 2001 11 Keeney M Chin Yee 1 We
35. TFSC FSC SSC FSC CD34 FSC FSC CD45 CD45 CD45dim CD34 5 5 2 CD34 CD34 CD34 CD45 CD34 CD45
36. 4 3 3 5 3 2 CD34 CD34 W 4 3 4 CA CRY CE NL JCCLS H1 A H2 P CD34 COD34 CD34
37. 5 3 3 2 Se Scaiier 0 0 400 B00 HOO 1000 Fnrwward Seaiter Gate G4 CB 002 R44 and RS CB O002 la Scaitisr 8 8 8 8 yy 0 10 0 2 400 BOD BO 1000 CDs FTC Farviad 5cgtar Gate No Gaite Gated Evwents 105832 Total Ewents 105832 Gate Events CD34 cells 93 CD45 cells 70683 beads 7074 dead cells 4476 5 3 2 beads
38. ir K et al Single platform flow cytometric absolute CD34 cell counts based on the ISHAGE guldelines Cytometry 34 61 70 1998 12 NCCLS Clinical Applications of Flow Cytometry Quality Assuranse and Immu nophenotyping of Lymphocytes Approved Guideline NCCLS Document H45 A NCCLS Wayne 1998
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