Detection of Human Hematopoietic Stem Cell Engraftment in the Livers of Adult Immunodeficient Mice by an Optimized Flow Cytometric Method

Overview

Journal Stem Cell Stud

Date 2011 May 24

PMID 21603093

Citations 16

Authors

Nicole L Varga

Alicia Barcena

Marina E Fomin

Marcus O Muench

Affiliations

Soon will be listed here.

Abstract

Immunodeficient NOD.Cg-Prkdc(scid) Il2rg(tm1Wjl)/SzJ (NSG) mice are a valuable resource to study human hematopoietic stem cells. Prolonged multilineage hematopoiesis indicates stem cell engraftment and generally is measured by flow cytometry. In this study, we took advantage of the multi-parameter detection afforded by modern flow cytometers to optimize detection of human hematopoiesis in NSG mice. Antigens widely expressed by mouse or human cells were evaluated as markers to distinguish mixtures of these cells to optimize and test the limits of chimerism detection. The bone marrow, spleen, and liver of NSG mice transplanted with human hematopoietic cells were analyzed for evidence of engraftment.Mouse bone marrow cells were best marked for exclusion by staining with a combination of CD45, TER-119, and anti-H-2K(d) monoclonal antibodies, whereas live human cells were most accurately identified by elimination of cell doublets and positive staining for CD59. Human stem cells (CD34(++)CD133(+)CD38(low)) and progenitors were detected in the bone marrow and liver, but not in the spleen. An unusual pattern of myeloid antigen expression was detected in the bone marrow and CD3(+)CD4(+)CD8(+) T-cells were detected in the spleen. We concluded that multicolor flow cytometric analysis that clearly distinguishes mouse and human cells offers accurate detection of human chimerism in NSG mice. Human hematopoiesis can be detected in the bone marrow and liver of NSG mice with T-lymphopoiesis, possibly occurring in the spleen.

Citing Articles

Fetal liver CD34 contain human immune and endothelial progenitors and mediate solid tumor rejection in NOG mice.

Celhar T, Li X, Zhao Y, Tay H, Lee A, Liew H Stem Cell Res Ther. 2024; 15(1):164.

PMID: 38853275 PMC: 11163708. DOI: 10.1186/s13287-024-03756-7.

A Monocytic Barrier to the Humanization of Immunodeficient Mice.

Du E, Muench M Curr Stem Cell Res Ther. 2023; 19(7):959-980.

PMID: 37859310 PMC: 10997744. DOI: 10.2174/011574888X263597231001164351.

A bioinspired and chemically defined alternative to dimethyl sulfoxide for the cryopreservation of human hematopoietic stem cells.

Gilfanova R, Callegari A, Childs A, Yang G, Luarca M, Gutierrez A Bone Marrow Transplant. 2021; 56(11):2644-2650.

PMID: 34155359 PMC: 8563414. DOI: 10.1038/s41409-021-01368-w.

Immunodeficient mice are better for modeling the transfusion of human blood components than wild-type mice.

Blessinger S, Tran J, Jackman R, Gilfanova R, Rittenhouse J, Gutierrez A PLoS One. 2020; 15(7):e0237106.

PMID: 32735605 PMC: 7394438. DOI: 10.1371/journal.pone.0237106.

Higher Serum Alanine Transaminase Levels in Male Urokinase-Type Plasminogen Activator-Transgenic Mice Are Associated With Improved Engraftment of Hepatocytes but not Liver Sinusoidal Endothelial Cells.

Fomin M, Beyer A, Publicover J, Lu K, Bakkour S, Simmons G Cell Med. 2017; 9(3):117-125.

PMID: 28713641 PMC: 5509021. DOI: 10.3727/215517916X693375.

References

Skubitz K, Campbell K, Skubitz A . CD66a, CD66b, CD66c, and CD66d each independently stimulate neutrophils. J Leukoc Biol. 1996; 60(1):106-17. DOI: 10.1002/jlb.60.1.106. View

Muench M, Cupp J, Polakoff J, Roncarolo M . Expression of CD33, CD38, and HLA-DR on CD34+ human fetal liver progenitors with a high proliferative potential. Blood. 1994; 83(11):3170-81. View

Xiao M, Dooley D . Cellular and molecular aspects of human CD34+ CD38- precursors: analysis of a primitive hematopoietic population. Leuk Lymphoma. 2000; 38(5-6):489-97. DOI: 10.3109/10428190009059267. View

Muench M, Suskind D, Barcena A . Isolation, growth and identification of colony-forming cells with erythroid, myeloid, dendritic cell and NK-cell potential from human fetal liver. Biol Proced Online. 2003; 4:10-23. PMC: 145552. DOI: 10.1251/bpo29. View

Schmidt M, Appel M, Giassi L, Greiner D, Shultz L, Woodland R . Human BLyS facilitates engraftment of human PBL derived B cells in immunodeficient mice. PLoS One. 2008; 3(9):e3192. PMC: 2527131. DOI: 10.1371/journal.pone.0003192. View

Hayakawa J, Hsieh M, Anderson D, Phang O, Uchida N, Washington K . The assessment of human erythroid output in NOD/SCID mice reconstituted with human hematopoietic stem cells. Cell Transplant. 2011; 19(11):1465-73. PMC: 3879801. DOI: 10.3727/096368910X314161. View

Yin A, Miraglia S, Zanjani E, Almeida-Porada G, Ogawa M, Leary A . AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood. 1998; 90(12):5002-12. View

Jersmann H . Time to abandon dogma: CD14 is expressed by non-myeloid lineage cells. Immunol Cell Biol. 2005; 83(5):462-7. DOI: 10.1111/j.1440-1711.2005.01370.x. View

DiGiusto D, Chen S, Combs J, Webb S, Namikawa R, Tsukamoto A . Human fetal bone marrow early progenitors for T, B, and myeloid cells are found exclusively in the population expressing high levels of CD34. Blood. 1994; 84(2):421-32. View

10.

Ziegler-Heitbrock L . The CD14+ CD16+ blood monocytes: their role in infection and inflammation. J Leukoc Biol. 2006; 81(3):584-92. DOI: 10.1189/jlb.0806510. View

11.

Kamel-Reid S, Dick J . Engraftment of immune-deficient mice with human hematopoietic stem cells. Science. 1988; 242(4886):1706-9. DOI: 10.1126/science.2904703. View

12.

Terstappen L, Huang S, Safford M, Lansdorp P, Loken M . Sequential generations of hematopoietic colonies derived from single nonlineage-committed CD34+CD38- progenitor cells. Blood. 1991; 77(6):1218-27. View

13.

Schipper L, van Hensbergen Y, Fibbe W, Brand A . A sensitive quantitative single-platform flow cytometry protocol to measure human platelets in mouse peripheral blood. Transfusion. 2007; 47(12):2305-14. DOI: 10.1111/j.1537-2995.2007.01472.x. View

14.

Murray L, Mandich D, Bruno E, DiGiusto R, FU W, Sutherland D . Fetal bone marrow CD34+CD41+ cells are enriched for multipotent hematopoietic progenitors, but not for pluripotent stem cells. Exp Hematol. 1996; 24(2):236-45. View

15.

Erlandsen S, Greet Bittermann A, White J, Leith A, Marko M . High-resolution CryoFESEM of individual cell adhesion molecules (CAMs) in the glycocalyx of human platelets: detection of P-selectin (CD62P), GPI-IX complex (CD42A/CD42B alpha,B beta), and integrin GPIIbIIIa (CD41/CD61) by immunogold labeling and.... J Histochem Cytochem. 2001; 49(7):809-19. DOI: 10.1177/002215540104900702. View

16.

Watkins N, Gusnanto A, de Bono B, De S, Miranda-Saavedra D, Hardie D . A HaemAtlas: characterizing gene expression in differentiated human blood cells. Blood. 2009; 113(19):e1-9. PMC: 2680378. DOI: 10.1182/blood-2008-06-162958. View

17.

Kyoizumi S, Baum C, KANESHIMA H, McCune J, Yee E, Namikawa R . Implantation and maintenance of functional human bone marrow in SCID-hu mice. Blood. 1992; 79(7):1704-11. View

18.

Macchiarini F, Manz M, Palucka A, Shultz L . Humanized mice: are we there yet?. J Exp Med. 2005; 202(10):1307-11. PMC: 2212979. DOI: 10.1084/jem.20051547. View

19.

Shultz L, Ishikawa F, Greiner D . Humanized mice in translational biomedical research. Nat Rev Immunol. 2007; 7(2):118-30. DOI: 10.1038/nri2017. View

20.

Neildez-Nguyen T, Wajcman H, Marden M, Bensidhoum M, Moncollin V, Giarratana M . Human erythroid cells produced ex vivo at large scale differentiate into red blood cells in vivo. Nat Biotechnol. 2002; 20(5):467-72. DOI: 10.1038/nbt0502-467. View