Non-human primates (NHPs) are valuable as preclinical resources that bridge the gap between basic science and clinical application. B cells from NHPs have been utilized for the development of B-cell targeted drugs and cell-based therapeutic modalities; however, few studies on the ex-vivo expansion of monkey B cells have been reported. In this study, we developed a highly efficient ex-vivo expansion protocol for monkey B cells resulting in 99% purity without the requirement for prior cell-enrichment procedures. To this end, monkey peripheral blood mononuclear cells (PBMCs) were stimulated for 12 days with cells constitutively expressing monkey CD40L in expansion medium optimized for specific and massive expansion of B cells. The B cells expansion rates obtained were 2-5 times higher than those previously reported in humans, with rates ranging from 7.9 to 16.6 fold increase. Moreover, expanded B cells sustained high expression of co-stimulatory molecules including CD83 and CD86 until day 12 of culture, and the simple application of a brief centrifugation resulted in a CD20+ B cell purity rate of greater than 99%. Furthermore, small amounts of CD3+CD20+BT-like cells were generated and CD16 was expressed at moderate levels on expanded B cells. Thus, the establishment of this protocol provides a method to produce quantities of homogeneous, mature B cells in numbers sufficient for the in vitro study of B cell immunity as well as for the development of B cell-diagnostic tools and cell-based therapeutic modalities.
|Number of pages
|Biochemical and biophysical research communications
|Published - 2016 Apr 22
Bibliographical noteFunding Information:
This work was supported by a grant from the Korea Healthcare Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), and funded by the Ministry for Health and Welfare, Republic of Korea (Grant No. HI13C0954 ). The authors would like to thank Jung WY, Kim MS, Lee GE, and Kim JE for caring for all the NHPs. We are also indebted to Nam HY and Lim HJ for their continuous support in this study.
© 2016 Elsevier Inc. All rights reserved.
- B cell
- CD40-CD40L signals
- Ex-vivo expansion
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology