I was looking for a cardiac muscle cell line and came across HL1 but have no idea about these cells. Any input would be a great help.
Thank you.
Need to contact William C. Claycomb at LSU School of Med.
http://www.medschool.lsuhsc.edu/biochemistry/faculty_detail.aspx?name=claycomb_william
We found them to be a bit inconsistent, and expensive. But from a cardiac standpoint your limited for alternatives. Can only order medium from sigma (51800C; $120/ 500mL), plus the fibronectin, FBS (special lot from sigma, 12103C; $300+/500mL). In addition you need NE, L-Glut, PS, Trypsin and Soybean Trypsin inhibitor.
To add to Chad's comments, there's H9c2 also, which can be induced/paced to beat and are rat ventricular. In contrast, HL-1 is mouse atrial and beats spontaneously. HL-1 basal medium and qualified lot of FBS are both from Sigma/SAFC and not cheap, plus norepinephrine.
The split has to be 1 to 3 and the next split occurs only at 100% confluence, when every few cells are seen beating spontaneously. They also need to be fed daily or 2x volume (10mL in T-25) for 2 days or weekend, and the medium needs to be kept in the dark, with the hood light off during work.
Thank you Chad and Gin,
This is valuable information to start with.
I have used H9C2 cells previously as cardiac-like cells (after differentiating them with all-trans retinoic acid) but never could induce them to beat unfortunately and when we tried to publish that data reviewers gave us hard time and we had to use rat neonatal primary cardiomyocytes along with H9C2.
You will find that you get the same criticism with the HL-1 cell line. At minimum you will have to prove the major findings in adult primary ventricular cardiomyocytes.
From our experience I can tell you it is not cheap to work with this cell line, you need to use the special lot of medium Gin and Chad mentioned. We tried to maintain the cells in DMEM, OPTIMEM or IMDM supplemented with components present in Claycomb medium and simply cells did not take off. The major aproximation we made was to feed cells in P100 dishes with Claycomb medium and once passed to 24, 12 or 6- well plates change medium to Optimem supplemented with 3 % FBS.
Hi,
Is it mandatory to use the special Fetal Bovine Serum, HL-1 Cell Screened, or standard FBS (Heat Inactivated) could be fine ?
Thanks a lot !
You need the correct medium (Claycomb) and an incubator...cells can be derived from any supplier William C. Claycomb at LSU School of Med. Most labs working on cardial biology use and have HL-1 cells
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