I got gene expression profiles from single cell sequencing. Some cells have high expression of mbp (mature oligodendrocyte), but low expression of olig1,2, sox10 and Mag (Oligodendrocyte lineage). Interestingly, these cell have moderate expression of NG2 (CSPG4), but no expression of PDGFRa (both are OPC marker). Could we judge a cell sole by the transcript profile? what are my cells? OPC, mature oligodendrocytes, or other possibilities? Thanks
Hello Lingyan,
From the profiles you mentioned, it seems like your cells are going to mature oligodendrocyte lineage thats why a full blown expression of mbp while moderate or no expression of OPC markers.
You can confirm it on protein level by choosing a marker of mature oligodendrocytes and OPC. Transcripts level should be confirmed by protein analysis by ICC or WB. Once you get same pattern in protein expression also, you can be sure that the cells are mature oligodendrocytes.
Hope it helps!
Hi,
I think your cells are mature oligodendrocytes.
Markers of premyelinating oligodendrocyte: NG2-, PDGRFalpha-, O4+, OLIG1+, OLIG2+, SOX10+, Galactocerebroside+, MAG-, MBP-
Markers of myelinating oligodendrocyte: NG2-, PDGFRalpha-, O4+, OLIG1+/-, OLIG2+/-, SOX10+, Galactocerebroside+, MAG+, MBP+
You should confirm your results with RT-qPCR and then at protein level ELISA or WB.
Good luck!
Hi Lingyan , I've done some single cell sequencing of neuronal lines - some cells showed neuron specific mRNA AND oligodenritic or astrocyte markers (and of course some showed mRNA for just one or the other).
I thought it was quite possible that the multiple/conflicting levels of these mRNAs might reflect that cell as possibly an early progenitor cell (that hasn't quite made its mind up which type to be). Do these cells also express pluripotency markers to some level (Oct4/nanog?)?
However it is always dangerous in interpreting data on a single cell as mRNA expression levels can occur in "bursts" and the amounts are "stochastic" at the single cell level. Also this is the level of that mRNA at the time of analysis (it may be just about to increase or decrease at the time of cell lysis). Really to make a judgement of which way a population is "going" from single cell data you need to examine a number of single cells (if memory serves me right Stahlberg et al 2013 (qPCR work flow paper) suggests that a minimum of 50 single cells need to be examined before conclusions can be reached.
So to answer your question "Could we judge a cell sole by the transcript profile? " the answer (according to current thinking) is "no" :)
Hope this helps,
Gary
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