Establishment of murine allogeneic umbilical cord blood cell transplantation model and study on the mechanism of keratinocyte growth factor enhancing immune reconstitution after transplantation..

Yi WANG; Guang-Hua CHEN; De-Pei WU; Hai-Wen HUANG; Ying WANG; Yang XU; Xiao MA; Ai-Ning SUN; Hui-Rong CHANG

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Establishment of murine allogeneic umbilical cord blood cell transplantation model and study on the mechanism of keratinocyte growth factor enhancing immune reconstitution after transplantation..

Author: Yi WANG ¹ ; Guang-Hua CHEN ; De-Pei WU ; Hai-Wen HUANG ; Ying WANG ; Yang XU ; Xiao MA ; Ai-Ning SUN ; Hui-Rong CHANG
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1. Jiangsu Institute of Hematology, Key laboratory of Thrombosis and Hemostasis, Ministry of Health, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Publication Type:Journal Article
MeSH: Animals; Cord Blood Stem Cell Transplantation; Fetal Blood; transplantation; Fibroblast Growth Factor 7; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Mice; Mice, Inbred BALB C
From: Chinese Journal of Hematology 2010;31(2):82-87
CountryChina
Language:Chinese
Abstract:
OBJECTIVETo explore the impact and mechanism of keratinocyte growth factor (KGF) on immune reconstitution post murine allogeneic umbilical cord blood transplantation (UCBT).
METHODSPerpheral blood (PB) from 19.5-day embryos post-conception (E 19.5 d) mice was used as umbilical cord blood (UCB) graft. Thirty-two BALB/c mice were randomly assigned to 4 groups with 8 mice each in the first cohort UCBT. Mice were infused with PBS (control group) or 1 x 10(6) (group 1A), 2 x 10(6) (group 1B), 3 x 10(6) UCB mononuclear cells (MNCs) (group 1C), respectively. Twenty-four BALB/c mice were randomly assigned to 3 groups with 8 mice each in the second cohort UCBT. Mice were injected with 1 x 10(6) (group 2A), 2 x 10(6) (group 1B) or 3 x 10(6) (UCB) MNCs (group 2C). All mice received platelet transfusion on +8d. Sixteen BALB/c mice were randomly assigned to 2 groups with 8 mice each in the third cohort UCBT. Mice were injected s.c. with KGF (group 3) or PBS (control group) before TBI. All mice were injected with 2 x 10(6) UCB MNCs and were supported with platelet transfusion on +8 d. The survival time, splenic lymphoid cell subsets, sjTREC assay were observed after UCBT.
RESULTSThe 100-day survival of mice were 2, 3 and 3 in group 1A, 1B, 1C and 7, 8, 8 in group 2A, 2B, 2C, respectively. The splenic T, NKT, NK and B cell counts on +35 d were (9.57 +/- 0.74) x 10(6), (0.64 +/- 0.06) x 10(6), (1.43 +/- 0.10) x 10(6) and (19.13 +/- 1.50) x 10(6) in control group, respectively; while were (13.47 +/- 0.74) x 10(6), (0.89 +/- 0.03) x 10(6), (1.79 +/- 0.04) x 10(6) and (20.50 +/- 0.91) x 10(6) in group 3, respectively, being significantly higher than in control group. The sjTREC level was 182.2 +/- 10.7copies per 10(5) cells in control group; while was 224.2 +/- 9.6 copied per 10(5) cells in group 3, being significantly higher than in control group (P = 0.019).
CONCLUSIONSPeripheral blood from E19.5d is rich in hematopoietic stem cells. A murine allogeneic UCBT model with platelet support on +8 d is established. KGF treatment can enhance thymic output and improve T cell immune reconstitution after UCBT.