Cryopreserved Human Bone Marrow Stroma is Fully Functional In vitro

Abstract

Human-marrow long-term culture (LTC) enables maintenance of both stromal and haemopoietic elements of normal bone marrow (NBM) in vitro for 4-6 months. Stroma-based cultures are critical for quantitation of long-term culture initiating cells (LTC-IC), the most primitive human haemopoietic cells measurable in vitro. Supply of NBM can be sporadic, and up to 3-4 weeks in culture is required for stromal maturity. Stroma availability for experimental purposes can therefore be limited. Efforts to produce transformed human and transfected murine stromal cell lines comparable to NBM stroma have had some success. As an alternative, we investigated cryopreserved NBM and cryopreserved performed stroma. Function of cryopreserved and control fresh NBM stroma was similar when evaluated for up to 12 weeks in LTC. We have also demonstrated that stroma derived from cryopreserved NBM or performed cryopreserved NBM stroma can sustain third-party haemopoiesis as efficiently as fresh NBM stroma in LTC. Batched cryopreserved stroma is a convenient, rapidly available, source of functional stroma which avoids the logistic difficulties and lack of standardization associated with stroma from fresh NBM. This important advance will enhance the use of stroma-based LTC in studies of human haemopoiesis.

Impact of age on the efficacy of bone marrow mononuclear cell transplantation in experimental stroke

“…The cells used in our study had been cryopreserved prior to use, while the majority of positive therapeutic results were obtained in studies using freshly prepared BM MNC. Cryopreservation does not seem to affect the hematopoietic potential [29]. However, it is unclear whether freezing and/or thawing alters potential neuroprotective capabilities, which probably depend on completely different physiological processes.…”

Mild Heat Shock Induces Proliferation, Alkaline Phosphatase Activity, and Mineralization in Human Bone Marrow Stromal Cells and Mg-63 Cells In Vitro

“…The confluent BMSCs were detached with Trypsin‐EDTA, washed, and stored in liquid nitrogen until use. Cryopreservation of human BMSCs has been shown to cause no significant in vitro functional change (18) . The osteoblast‐like MG‐63 cell line was cultured in DMEM with 10% FCS and 1 mM L ‐glutamine and passaged every 3–4 days.…”

Mesenchymal Stem Cells for Regenerative Therapy: Optimization of Cell Preparation Protocols

“…This allows for great flexibility in the clinical setting, but extreme caution is needed on possible adverse effects on MSCs. Although there are many preclinical studies showing that cryopreservation does not change the biological behavior of MSCs such as differentiation, growth, and/or surface marker expression [47, 48], on the other hand, there are reports warning hazardous effects by cryopreservation [49]. Further refinement of the protocol is warrantied.…”