Obtaining Mesenchymal Stem Cells from the Femoral Condyle During ACL Reconstruction: A Pilot Study
E.A.G. Krieger, A. Pereira, F.C. Karam, D.R. MarinowicObjective:
To evaluate the feasibility of obtaining mesenchymal stem cells (MSCs) from the lateral femoral condyle during anterior cruciate ligament (ACL) reconstruction surgeries, using different needles and cell separation methods.
Methods:
This prospective experimental pilot study included 17 patients undergoing ACL reconstruction. In the first phase, two bone marrow tissue samples were collected from each participant using a 16G epidural needle and a bone marrow biopsy needle, with 10 mL per sample. Samples were processed using three protocols: centrifugation, decantation, and density gradient separation. The material obtained was cultured in supplemented DMEM medium, monitored by optical microscopy, and evaluated for cell adherence, morphology, and colony formation over 30 days. In the second phase, samples from an additional 12 participants were processed by decantation and centrifugation using the same protocols. Four samples were also analyzed by flow cytometry.
Results:
No differences were observed in volume or visual appearance between samples obtained with the two needle types. The density gradient method produced a translucent supernatant, whereas centrifugation and decantation resulted in cellular debris. After seven days, all samples demonstrated colony-forming units. At 30 days, cultures showed greater than 80% confluence of adherent, spindle-shaped cells consistent with MSC morphology. The four samples analyzed by flow cytometry showed results compatible with mesenchymal stem cells.
Conclusion:
The acquisition of MSCs from the lateral femoral condyle is feasible using a 16G epidural needle and separation methods, such as decantation, without compromising cell quality. The technique is low cost, safe, and can be performed during knee procedures.