Hi there! I've decided to work with reagents from stemcell, namely SFEM II & erythroid expainsion supplement.
The purpose is to expand erythroblasts from freshly isolated PBMCs for somatic cell reprogramming via sendai virus.
All the protocols I've gone through multiple times say the same statement "PBMCs were cultured for 7 days till the day of viral transduction in XX media supplemented with YY" and so on.
The problem is, I do not know whether I am expected to culture those cells on 6-well plate format that is tissue-culture treated, or should I go for plates that we use for embryoid body generation that are low-attachment.
Thanks in advance!
I have not worked with your erythrocyte progenitors but have cultured PBMCs. There are some protocols(specifically generation of LCLs and some antigen specific T cell generation methods) where you may not change media for 7 days so when they say "cultured for 7 days" they may mean literally chuck it in the incubator and forget about it for 7 days(still check for contamination etc). The two methods from stem cell tech linked below both have media change steps though so you won't get off so easy.
The following two stem cell tech bulletin/manuals could be used as reference. In particular section 5.0 in "Integrated Workflow for the Isolation, Expansion, and Reprogramming of Erythroid Progenitor Cells" seems to cover using that media system for your purposes. It actually has centrifugation steps to change media which is not common in many primary cell PBMC culture methods. Typically media change is done by dilution or allowing cells to settle and gently removing 80-90% of the media(see the first media change in the technical bulletin for SFEM II media). I guess your cells are hardy enough. It looks like they use TC coated 6 well plates but you could probably also use uncoated 6 well plates to the same effect(just for the progenitor expansion steps). When you shift to your reprograming methods just replate the cells on your desired dish.
https://www.stemcell.com/stemspan-media-and-supplements-for-the-expansion-and-differentiation-of-erythroid-progenitor-cells-lp.html
https://cdn.stemcell.com/media/files/manual/MA29795-Integrated_Workflow_Isolation_Expansion_Reprogramming_Erythroid_Progenitor_Cells_2_1.pdf?_ga=2.84777567.751598883.1578009149-1816317027.1578009149
For a peer review citation this one uses the above protocol:Article Generation of induced pluripotent stem cells (iPSCs) from a ...
I think that covers your specific questions. As a reminder I've never worked with your cells so if you need specific advice for your cells reach out to stem cell tech. You're already using an expensive kit from them so reach out, they want you to succeed(so you'll keep buying kits)!
I appreciate the answer very much, that's all I needed to hear indeed. Thank you so much.