I am trying to study B cell maturation in vitro, and have found some (older) literature which shows people have done this by putting CD34+ cells from cord blood onto human bone marrow stromal cells (Comparative studies of different stromal cell microenvironments in support of human B-cell development).
However, I would like to have a cell line instead of having to buy and isolate bone marrow stromal cells. Does anyone know of a HUMAN cell line that supports B cell development?
I think the best way would be you isolate the stromal cells from the bone marrow, because it's closer the natural process of maturation, and the best cell to use, would be the CD34 from bone marrow too. But I know that is not easy to obtain bone marrow to do that. So, one option that you can try, and the protocol for isolation is easy, it's isolate the stromal cell from the cord blood, so you can expand for some passages, and freeze some aliquots to do your experiment. You can use the same cord blood that you will isolate the CD34 and isolate the stromal cells. We have a stromal cell line, but that cell lines is not good to work because those cells have a different behavior from the primary cells because they are modified genetically, for you see those cells are tumorigenic in mice.
Hi Hans:
I agree with Ricardo, if you can expand primary BMSCs that would be much better. You can also use HS-5 cell line, a fibrobalstoid line (HPV-transformed) from normal human bone marrow. You can obtain it from ATCC, I think it has the potential to drive normal hematopoiesis (not really sure). I have used it many times to explore its interaction with malignant plasma cells.
Good luck
This is really good to know! Ricardo, Jahangir, thank you! Just a couple more questions if you guys would:
It would be easier for me to get cord blood than bone marrow. I didn't know cord blood had a stromal cell population. Do you know of a protocol for isolating stromal cells from cord blood? Is it just like bone marrow (plate them and the cells that attach are the stromal cells)? And if I used these, would it be the similar to using bone marrow stromal cells? From what I've read, embryonic and adult hematopoiesis differ somewhat.
Do the CD34 HSCs and the bone marrow stromal cells have to come from the same donor?
Sorry, that was a little more than a couple questions :)
Cord tissue cannot be an "anatomic" site for hematopoiesis, it's a source for collection of HSCs and MSCs due to the high cell traffic. Yolk sac, liver, lymph nodes during embryogenesis and BM (and for a short time liver) after birth are predominant hematopoietic tissues, so you should look for BMSCs or stromal cells from lymph nodes to investigate lymphopoiesis, still you may also be able to isolate stromal cells from cord tissue but is difficult to ascertain whether they have the same functions as in the real microenvironment of the BM, and yes generally neonatal hematopoiesis differs a lot from adult hematopoiesis.
I see. Thank you, Jahangir.
It looks like the easiest route I have is to isolate BM stromal cells and CD34 HSC from BM to study B cell maturation. I could use CD34 HSCs from cord blood as well though, correct?
Yes!, that's right. If your plan were on mice models the life would be much easier, good luck with your endeavor anyways!
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