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Published on: February 16, 2024
Ex vivo expanded autologous hematopoietic stem cells stimulate improved immunogenic activity: an in-vitro preliminary
Ramesh Thevendran1,2, Yek Song Quek3, Sherlyn Low Si Tong3
1Faculty of Applied Science, AIMST University, Jalan Bukit Air Nasi, Semeling, 08100, Bedong, Kedah, Malaysia. rthevendran@yahoo.com.
Background:
Hematopoietic stem cell therapy is a cornerstone of regenerative medicine for treating haematological and immunological disorders. While autologous HSCs avoid the complications of graft-versus-host disease, current expansion protocols often rely on expensive synthetic media and xeno-derived components that hinder clinical translation. This study evaluates a minimalist, "in-house" expansion protocol designed for cost-effectiveness and complete human and Malaysian cGMP process compatibility.
Methods:
CD34 + cells were isolated from the peripheral blood of healthy donors and expanded using the in-house methodology consisting of autologous plasma supplemented with stem cell factor and interleukin-3. The expanded HSCs were co-cultured with autologous peripheral blood mononuclear cells to assess their stimulatory potential. Parameters measured included cell viability, cell purity, total white blood cell expansion event counts via flow cytometry, and Interferon-gamma secretion via ELISA.
Results:
The expansion protocol achieved good results, with viable CD34 + purity and a marginal CD71 + percentage. Functional assays revealed that the presence of expanded HSCs exerted a potent stimulatory effect on the immune environment. The co-culture group containing expanded HSCs and corresponding donor PBMC demonstrated a strong increase in total WBC event counts, reaching around average 3000 counts compared to counts < 500 and even less in the control group. Furthermore, this interaction induced strong immunogenic activity, characterised by high levels of IFN-gamma secretion, whereas the control groups showed negligible cytokine production.
Conclusions:
These findings demonstrate that ex vivo expanded autologous HSCs act as a strong biological stimulant, significantly aiding leukocyte proliferation and immune signalling. Our expansion methodology provides a safe, human-compatible, and effective framework for good improvement of such haematological factors. These results further corroborate the potential for this approach in future clinical therapies aimed at boosting immune recovery, overall patient wellness and haematological parameters.
