Effects of supravital fluorochromes used to analyze the in vivo homing of murine lymphocytes on cellular function

Samlowski, Wolfram E.; Robertson, Bekkie A.; Draper, Bradley K.; Prystas, Elizabeth; McGregor, John R.

doi:10.1016/0022-1759(91)90236-9

Cited by 88 publications

(63 citation statements)

References 19 publications

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“…Previous studies have demonstrated that PKH26 staining does not alter cell function, [13][14][15][16] and our results support this conclusion given that PKH26-labeled cells had the same long-term repopulating activity as unstained cells (data not shown). After implantation, the donor cells were identified by PKH26 labeling and by antibody staining using FACS analysis.…”

Section: Implantation Of Donor Cellssupporting

confidence: 90%

Murine hematopoietic stem cell distribution and proliferation in ablated and nonablated bone marrow transplantation

Zhong

Zhan

Anderson

et al. 2002

Blood

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The engraftment of donor bone marrow (BM) cells in nonablated mice is inefficient. Niche availability has been thought to be the reason, and cytoablation with irradiation or cytotoxic agents is routinely used with the belief that this frees the preoccupied niches in recipients. In this study, donor cell redistribution and proliferation in ablated and nonablated mice were compared by implanting donor cells directly into the femur cavity of

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Section: Implantation Of Donor Cellssupporting

confidence: 90%

Murine hematopoietic stem cell distribution and proliferation in ablated and nonablated bone marrow transplantation

Zhong

Zhan

Anderson

et al. 2002

Blood

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“…Their ability to differentiate is governed primarily by their surrounding environment [19,41]. Conditions favorable for inducing MSCs to undergo osteogenesis, chondrogenesis, and adipogenesis are well established [31]; however, no study thus far has reported the induction of MSCs to differentiate into NPCs or annulus fibrosus cells. Increasing evidence has suggested some stem cell populations exhibit a remarkable degree of plasticity after they are transplanted in recipients [11,33,37,39,41,45].…”

Section: Discussionmentioning

confidence: 99%

“…The lipophilic dye PKH is a nonradioactive and noncytotoxic substance and possesses a fluorescence half-life of more than 100 days in erythrocytes [31] or MSCs [39]. The fluorescence emitted by cells attributable to PKH uptake is not transferred to the neighboring cells but is transferred to the daughter cells during division [31,32,38]. Thus, the double-stained cells observed in the coculture may have been generated by spontaneous cell fusion of the MSCs (red fluorescent marker) and NPCs (green fluorescent marker).…”

Section: Discussionmentioning

confidence: 99%

Mesenchymal Stem Cell and Nucleus Pulposus Cell Coculture Modulates Cell Profile

Niu

Yuan

Lin

et al. 2008

Clin Orthop Relat Res

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Spontaneous cell fusion can occur in cocultured stem cells. We examined whether telomerase activity change and cell fusion occurred in mesenchymal stem cell (MSC) and nucleus pulposus cell (NPC) coculture. MSCs and NPCs were labeled with PKH26 and PKH67 dyes and cocultured at a 50:50 ratio. An equal number of MSCs or NPCs were used as the control. After 14 days, cells were evaluated by cell growth, telomerase activity, quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry, and histologic observation. Cell fusion was confirmed by microscopic observation and fluorescence-activated cell sorter (FACS) analysis. The results suggested cell growth rate and telomerase activity were higher in cocultured cells than in NPCs cultured alone. The mRNA expression levels of the Type II collagen and aggrecan were elevated in cocultured cells. Immunohistochemical analysis revealed positive staining for Type II collagen and keratan sulfate in NPCs cultured alone and in a proportion of cocultured cells. Histologic observation revealed binucleated cocultured cells expressed both PKH dyes in the same location and slide focus. The FACS analysis revealed 42% of cocultured cells were doublestained. Cocultured cells partially maintained the NPC phenotype. The partially maintained phenotype of the NPCs may be attributable to spontaneous cell fusion in association with increased telomerase activity.

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“…PKH-26, a non-permeant fluorescent lipophilic probe that binds to cellular membrane, has been used to label several types of cells (Samlowski et al 1991, Ward et al 1993. Macrophages were washed three times in PBS and incubated with PKH26 (100 µg/ml) in PBS for 30 s at 4ºC.…”

Section: Formation Of the Parasitophorous Vacuolementioning

confidence: 99%

Internalization of components of the host cell plasma membrane during infection by Trypanosoma cruzi

Carvalho

Souza

Coimbra

1999

Mem. Inst. Oswaldo Cruz

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Effects of supravital fluorochromes used to analyze the in vivo homing of murine lymphocytes on cellular function

Cited by 88 publications

References 19 publications

Murine hematopoietic stem cell distribution and proliferation in ablated and nonablated bone marrow transplantation

Murine hematopoietic stem cell distribution and proliferation in ablated and nonablated bone marrow transplantation

Mesenchymal Stem Cell and Nucleus Pulposus Cell Coculture Modulates Cell Profile

Internalization of components of the host cell plasma membrane during infection by Trypanosoma cruzi

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