human HSC can been transplanted in mice(NSG,NOG,NOD/SCID) through caudal vein or right femur (RF). Chimeric rate in PBMC and BM, which is higher? Can different transplant ways cause difference of chimeric rate in different tissue? (right femur (RF) ---more chimeric rate in BM, caudal vein----more in PBMC? )
BM engraftment is higher than other tissues in most circumstances. Given enough time, Hematopoiesis distributes among the various hematopoietic tissues from the point of origen.
@Marcus O. Muench You mean with different transplantation protocol（caudal vein or right femur (RF)），and different check time， BM is always higher？
I found a paper "A synthetic three-dimensional niche system facilitates generation of functional hematopoietic cells from human-induced pluripotent stem cells"， its PB is higher than BM，That is why I am confused.
My lab's experience is that the blood has the lowest levels of chimerism. We have a paper coming out in the near future that I can post when available. We use NSG, not NOD/SCID treated as the authors describe, so that may make some difference. Perhaps more importantly, we isolate PBMC – not total leukocytes by red cell lysis, in our studies.
I'm a little puzzled by the CD34 staining in the BM in the figure. A discrete CD34bright population with almost no CD34low cells does not suggest active hematopoiesis. I don't know what to make of those findings.
Hyde,
Our paper came online. Fig.1 has the comparison of total human chimerism in different tissues.
Article Comparison of Human Hematopoietic Reconstitution in Differen...
1). There is no recovery of immunity with age in NSG mice, they are not 'leakey'. We use adults for transplants as it is more convenient. Tail vein injections are relatively easy and consistent in adults. Others have used neonatal transplants with succcess, but it does require timing births to transplants which is more complicated.
2) I'm not sure of the effects of the two different X-rays. Nonetheless, it is always good to perform a titrationnof does under your specific conditions. Multiple factors play into determining a subleathal dose.
Thanks so much.
I want to build this human to mice system from zero (My mice to mice transplantation is ok). And the failure almost kill me. I do what I can to make it possible. Can you give me some advice.
1.change the mice from NOD/SCID to NCG (Chinese NSG, I am not sure the quality of the mice).
2.I can only get peripheral blood hematopoietic stem cells mobilization with G-CSF in healthy donors. CB and fetal material are hard to get here.
3.I do bone marrow aspiration in live mice. BM is higher than PBMC, as you data showed. I do not want to miss positive cells, even 0.1%.
4.I have different injection skills (Caudal vein and intra-bone marrow injection)
5.I checked the x-ray again and again. ( the machine is unsteadily) .
Without success makes me doubt everything.
Hyde,
My guess is the adult blood HSC is your problem. If you looked in the supplemental data of our paper, we had almost no chimerism in NSG mice with normal adult blood HSC. We had some myeloid engraftment in NSG-3GS mice, but still no clear multilineage hematopoiesis. I have not tried mobilized PB-HSC, but the higher cell dose available may help. Still, in my experience even cord blood is not as good as the fetal tissue, even though they differ in age by as little as 20 weeks. My advice is to figure out a way to get cord blood.
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