Human brain endothelial cells (HUBEC) promote SCID repopulating cell expansion through direct contact

Xiangfei Cheng; Thomas Macvittie; Barry Meisenberg; Emily Welty; Ann Farese; Douglas Tadaki; Naoko Takebe

doi:10.1080/08977190701671662

Human brain endothelial cells (HUBEC) promote SCID repopulating cell expansion through direct contact

Xiangfei Cheng, Thomas Macvittie, Barry Meisenberg, Emily Welty, Ann Farese, Douglas Tadaki, Naoko Takebe

School of Medicine

Research output: Contribution to journal › Article › peer-review

Abstract

The objective of this study was to re-evaluate the previously published hematopoietic stem cell (HSC) expansion work using human brain endothelial cells (HUBEC). The expansion effect of contact and non-contact conditions was reported to be equivalent by others. However, we report here different results that the expansion can be achieved only with direct contact. We co-cultured human CD34+ cells with and without HUBEC contact for seven days with cytokines and the readouts were CD34+/CD38- phenotype and SCID repopulating cell (SRC) frequency. Also tested was the inhibitory effect of Wnt receptor inhibitor Dkk-1 on HUBEC contact ex vivo expansion; whether an increased expression of Wnt3 occurs on the HUBEC surface; and detection of an increased nuclear localization of beta-catenin in CD34+/CD38- cells in HUBEC contact culture condition. We conclude that the successful expansion by HUBEC contact culture is a candidate explanation based on the Wnt family protein, possibly Wnt3, expression on HUBEC.

Original language	English (US)
Pages (from-to)	141-150
Number of pages	10
Journal	Growth Factors
Volume	25
Issue number	3
DOIs	https://doi.org/10.1080/08977190701671662
State	Published - Jun 2007

Keywords

CD34+/CD38-
Contact culture
Cord blood CD34+ cells
Wnt/β-catenin

ASJC Scopus subject areas

Endocrinology
Clinical Biochemistry
Cell Biology

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10.1080/08977190701671662

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title = "Human brain endothelial cells (HUBEC) promote SCID repopulating cell expansion through direct contact",

abstract = "The objective of this study was to re-evaluate the previously published hematopoietic stem cell (HSC) expansion work using human brain endothelial cells (HUBEC). The expansion effect of contact and non-contact conditions was reported to be equivalent by others. However, we report here different results that the expansion can be achieved only with direct contact. We co-cultured human CD34+ cells with and without HUBEC contact for seven days with cytokines and the readouts were CD34+/CD38- phenotype and SCID repopulating cell (SRC) frequency. Also tested was the inhibitory effect of Wnt receptor inhibitor Dkk-1 on HUBEC contact ex vivo expansion; whether an increased expression of Wnt3 occurs on the HUBEC surface; and detection of an increased nuclear localization of beta-catenin in CD34+/CD38- cells in HUBEC contact culture condition. We conclude that the successful expansion by HUBEC contact culture is a candidate explanation based on the Wnt family protein, possibly Wnt3, expression on HUBEC.",

keywords = "CD34+/CD38-, Contact culture, Cord blood CD34+ cells, Wnt/β-catenin",

author = "Xiangfei Cheng and Thomas Macvittie and Barry Meisenberg and Emily Welty and Ann Farese and Douglas Tadaki and Naoko Takebe",

note = "Funding Information: We acknowledge the contribution of Dr William Morgan{\textquoteright} laboratory at the Department of Radiation Oncology at the University of Maryland in providing technical assistance with fluorescence microscopy analysis. We acknowledge Dr Ludy Dobrila and Dr Pablo Rubinstein at the New York Blood Center for providing CB cells and manuscript review. Supported by the Department of the Navy N3239802MP00102 (TM) and N000140610083 (NT).",

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doi = "10.1080/08977190701671662",

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AU - Cheng, Xiangfei

AU - Macvittie, Thomas

AU - Meisenberg, Barry

AU - Welty, Emily

AU - Farese, Ann

AU - Tadaki, Douglas

AU - Takebe, Naoko

N1 - Funding Information: We acknowledge the contribution of Dr William Morgan’ laboratory at the Department of Radiation Oncology at the University of Maryland in providing technical assistance with fluorescence microscopy analysis. We acknowledge Dr Ludy Dobrila and Dr Pablo Rubinstein at the New York Blood Center for providing CB cells and manuscript review. Supported by the Department of the Navy N3239802MP00102 (TM) and N000140610083 (NT).

PY - 2007/6

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N2 - The objective of this study was to re-evaluate the previously published hematopoietic stem cell (HSC) expansion work using human brain endothelial cells (HUBEC). The expansion effect of contact and non-contact conditions was reported to be equivalent by others. However, we report here different results that the expansion can be achieved only with direct contact. We co-cultured human CD34+ cells with and without HUBEC contact for seven days with cytokines and the readouts were CD34+/CD38- phenotype and SCID repopulating cell (SRC) frequency. Also tested was the inhibitory effect of Wnt receptor inhibitor Dkk-1 on HUBEC contact ex vivo expansion; whether an increased expression of Wnt3 occurs on the HUBEC surface; and detection of an increased nuclear localization of beta-catenin in CD34+/CD38- cells in HUBEC contact culture condition. We conclude that the successful expansion by HUBEC contact culture is a candidate explanation based on the Wnt family protein, possibly Wnt3, expression on HUBEC.

AB - The objective of this study was to re-evaluate the previously published hematopoietic stem cell (HSC) expansion work using human brain endothelial cells (HUBEC). The expansion effect of contact and non-contact conditions was reported to be equivalent by others. However, we report here different results that the expansion can be achieved only with direct contact. We co-cultured human CD34+ cells with and without HUBEC contact for seven days with cytokines and the readouts were CD34+/CD38- phenotype and SCID repopulating cell (SRC) frequency. Also tested was the inhibitory effect of Wnt receptor inhibitor Dkk-1 on HUBEC contact ex vivo expansion; whether an increased expression of Wnt3 occurs on the HUBEC surface; and detection of an increased nuclear localization of beta-catenin in CD34+/CD38- cells in HUBEC contact culture condition. We conclude that the successful expansion by HUBEC contact culture is a candidate explanation based on the Wnt family protein, possibly Wnt3, expression on HUBEC.

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Human brain endothelial cells (HUBEC) promote SCID repopulating cell expansion through direct contact

Abstract

Keywords

ASJC Scopus subject areas

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