Hemovac blood after total knee arthroplasty as a source of stem cells

Seon Ae Kim; Ho Youn Park; Yong-Woon Shin; Eun Jeong Go; Young Ju Kim; Yoo Chang Kim; Asode Ananthram Shetty; Seok Jung Kim

doi:10.21037/atm-20-2215

Hemovac blood after total knee arthroplasty as a source of stem cells

Ann Transl Med. 2020 Nov;8(21):1406. doi: 10.21037/atm-20-2215.

Authors

Seon Ae Kim¹, Ho Youn Park¹, Yong-Woon Shin², Eun Jeong Go¹, Young Ju Kim³, Yoo Chang Kim¹, Asode Ananthram Shetty⁴, Seok Jung Kim¹

Affiliations

¹ Department of Orthopedic Surgery, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
² Department of Orthopaedic Surgery, College of Medicine, The Inje University of Korea, Seoul, Republic of Korea.
³ Department of Nursing Education & Administration, Uijeongbu St. Mary's Hospital, The Catholic University of Korea, Seoul, Republic of Korea.
⁴ Canterbury Christ Church University, Faculty of Health and Wellbeing, Chatham Maritime, Kent, UK.

Abstract

Background: With increasing life expectancy, stem cell therapy is receiving increasing attention. However, its application is restricted by ethical concerns. Hence a need exists for design of safe procedures for stem cell procurement. Here, we investigated whether hemovac blood (HVB) is an appropriate stem cell source.

Methods: HVB concentrates (HVBCs) from 20 total knee arthroplasty (TKA) patients and bone marrow aspirate (BMA) concentrates (BMACs) from 15 patients who underwent knee cartilage repair were comparatively evaluated. A bone marrow aspiration needle was inserted into the anterior superior iliac spine. Aspiration was performed using a 50-mL syringe, including 4 mL of anticoagulant, followed by centrifugation to obtain BMACs. To obtain HVBCs, blood was aspirated from the hemovac immediately after TKA surgery. Different cell types were enumerated. Isolation of BMA and HVB mononuclear cells was performed using density gradient centrifugation. Non-hematopoietic fibroblast colonies were quantified by colony forming unit-fibroblast assay surface marker analysis of HVB, HVBC, BMA, and BMAC was performed via flow cytometry. Mesenchymal stem cells (MSCs) isolated from HVBCs and BMACs were examined for osteogenic, adipogenic, and chondrogenic differentiation potential. Gene expression analysis was performed by quantitative real-time polymerase chain reaction (qRT-PCR).

Results: The number of cells from HVB and HVBC was significantly lower than from BMA and BMAC; however, the number of colonies in HVBC and BMAC did not differ significantly (P>0.05). Isolated cells from both sources had a fibroblast-like appearance, adhered to culture flasks, and formed colonies. Under different culture conditions, MSC-specific surface markers (CD29, CD44, CD90, CD105), osteogenic markers [RUNX2, osteopontin, osteocalcin, and alkaline phosphatase (ALP)] and adipogenic markers (PPARγ and C/EBPα) were expressed. Moreover, SOX9, type II collagen, and aggrecan were significantly upregulated upon chondrogenic differentiation.

Conclusions: HVB from TKA patients is a useful source of stem cells for research.

Keywords: Bone marrow; bone marrow aspirate concentrate (BMAC); hemovac blood (HVB); stem cell; total knee arthroplasty (TKA).