CN105039158A

CN105039158A - Simple small bioreactor

Info

Publication number: CN105039158A
Application number: CN201510464421.4A
Authority: CN
Inventors: 金梦瑶; 祁轶之; 祁小平
Original assignee: Wuxi Sunlight Vale Life Technologies Co Ltd
Current assignee: Wuxi Sunlight Vale Life Technologies Co Ltd
Priority date: 2015-07-31
Filing date: 2015-07-31
Publication date: 2015-11-11

Abstract

The invention discloses a bioreactor designed through transformation based on a common 50 ml centrifugal tube in a biology laboratory. The reactor adopts the scheme that a section of thread in a tube cover of the original centrifugal tube is removed to form a potential channel; a small part of a thread at the lower part of the left end of the channel is also removed to form a notch; another notch is formed in a thread at the upper part of the right end of the channel, and a hole communicated with the inside of the tube is formed in the part, corresponding to the notch, of the tube wall. Through the adoption of the transformed design, inside gas and outside gas can be exchanged through the channel; by virtue of certain length and bendability of the channel, cultures in the tube can be prevented from being polluted by microbes outside the tube within the culture period of more than 7 days. When the simple small bioreactor is used for suspended cell culture, the cell density in the bioreactor is remarkably higher than that in the centrifugal tube completely sealed by the tube cover, so that the bioreactor can be used for research of multiple experiment parameters economically and conveniently, and is particularly suitable for cell culture media development and preliminary production technology development.

Description

A kind of easy small-sized biological reactor

Technical field

The invention belongs to bioengineering field, be specifically related to a kind of easy small-sized biological reactor.

Background technology

The production of human or animal's vaccine and the production of portion gene engineering medicine generally involve large-scale cell cultures, grow well to make cell, need to provide suitable growth conditions to cell, comprising nutritive ingredient that is abundant and balance, stable temperature, osmotic pressure, pH, suitable oxygen and carbon dioxide dividing potential drop etc.

The culture conditions equipment that can offer the best at present is the bio-reactor accurately to be controlled various kinds of cell growth important parameter by computer.The maximum shortcoming of this equipment is expensive, complicated operation, can the limited amount of parallel test simultaneously.In the process of process exploitation, especially usually need to observe many parameters and condition in the selection and performance history of substratum simultaneously, such as several, tens even up to a hundred, adopt aforesaid device obviously unrealistic in this case.

In order to overcome above-mentioned shortcoming, people manage to have developed various other forms of small-sized bio-reactor, as 96 hole depth orifice plates, and nonrecoverable blender jar, wave bio-reactor, the 50mL bio-reactor etc. of band air-permeable envelope.

Summary of the invention

Goal of the invention: the object of the invention is to for the deficiencies in the prior art, provides a kind of with low cost, easy to operate, can carry out the easy small-sized biological reactor of a large amount of parallel experiment.

Technical scheme: in order to reach foregoing invention object, the present invention has specifically come like this: a kind of easy small-sized biological reactor, comprise and there is externally threaded body and the female pipe lid of tool, passage is there is between described body and pipe lid, inside and outside body, gas can exchange, with this solve closed container for during suspension cell high-density culture because hypoventilation is to the restriction of stand density.

The one of described passage forms scheme and is: pipe lid internal thread is removed one section and formed passage one, the internal thread below passage one is removed one and is cut into breach one; Corresponding breach one place of body outside screw also removes one section and forms breach two, and the outside screw above respective channel one place has through hole, and breach one-breach two-passage one-through hole forms passage.

Wherein, be provided with corresponding projection one place above one group of parallel projection one, pipe lid internal thread above body outside screw and be provided with one group of parallel projection two, when pipe lid and body rotate screw up time, projection one and projection two can engage mutually; The number of described projection one, projection two is between 1 ~ 5.

Beneficial effect: when the present invention is used for suspension cell culture, cell density is significantly higher than with the complete totally enclosed centrifuge tube of pipe lid, economy groping of multiple experiment parameter be can carry out easily, cell culture media formulations exploitation and the exploitation of preliminary production technique are particularly suitable for.

Accompanying drawing explanation

Fig. 1 is pipe structure schematic diagram of the present invention;

Fig. 2 is pipe lid structural representation of the present invention.

Embodiment

Embodiment 1:

The easy small-sized biological reactor of one as shown in Fig. 1 ~ 2, comprise the body 1 with outside screw 3 and the pipe lid 2 with internal thread 4, described pipe lid internal thread is removed one section and is formed passage 1, the internal thread below passage 1 is removed one and is cut into breach 1; Body outside screw 3 corresponding breach 1 place also removes one section and forms breach 27, and the outside screw above respective channel 1 place has through hole 8, and breach one 6-breach two 7-passage one 5-through hole 8 forms passage; Wherein, further improve, be provided with corresponding projection 1 place above one group of parallel projection 1, pipe lid internal thread above body outside screw 3 and be provided with one group of parallel projection 2 10, when pipe lid 2 and body 1 rotate screw up time, projection 1 and projection 2 10 can engage mutually, and the number of described projection 1, projection 2 10 is 3.

Embodiment 2:

Cultured continuously is on the impact of substratum sterile state in holding tube: often prop up in bio-reactor and add 10mlDMEM nutrient solution respectively, totally 10.Put 37 DEG C of shaking tables (ThermoMAXQ420HP), speed 200rpm cultivates, and in every day observation tube, substratum color and turbidity change, judge whether nutrient solution is contaminated by bacterial.Continuous Observation 7 days, does not find that any nutrient solution has pollution sign.

Embodiment 3:

Cultured continuously is on the impact of substratum steam output change in pipe: often prop up in bio-reactor and add 10mlDMEM nutrient solution respectively, totally 5.Put 37 DEG C of shaking tables (ThermoMAXQ420HP), speed 200rpm cultivates, and the method for to weigh every day with electronic balance, in observation tube, the change of liquid evaporation amount, the results are shown in Table 2.Result shows often to prop up steam output every day of substratum in bio-reactor at about 20-30ul, and long-term cultivation can't cause the excessive variation of culture volume and osmotic pressure because of evaporation.

Table 1

Embodiment 4:

Get common centrifuge tube and each 5 of the present invention, adding initial density is respectively 5 × 10 ⁵the BHK-21 cell 10ml of/ml, substratum is the low serum suspension medium of BHK-21 cell that Wuxi sunlight paddy Life Science company limited produces.Put shaking table and carry out suspension culture (ThermoMax2000), CO put into by shaking table ₂incubator (Thermo1300 series A 2), culture condition is set to 37 DEG C, CO ₂concentration is 5%, saturated humidity.The culture that took a morsel respectively in 24,48 hours carries out cell counting, the results are shown in Table 2, and surveys pH value with pH meter (SartoriusPB-11), the results are shown in Table 3.

Result shows that cultured cells density of the present invention is all significantly higher than common centrifuge tube at different time, and the pH value after 48 hours simultaneously in the present invention still maintains in applicable scope, and common centrifuge tube exceed applicable scope.

Table 2

Table 3

Claims

1. an easy small-sized biological reactor, comprise the body (1) with outside screw (3) and the pipe lid (2) with internal thread (4), it is characterized in that, there is passage between described body (1) and pipe lid (2), the inside and outside gas of body (1) can exchange.

2. bio-reactor according to claim 1, is characterized in that, described pipe lid internal thread is removed one section and formed passage one (5), the internal thread of passage one (5) below is removed one and is cut into breach one (6); Body outside screw (3) corresponding breach one (6) place also removes one section and forms breach two (7), outside screw above respective channel one (6) place has through hole (8), and breach one (6)-breach two (7)-passage one (5)-through hole (8) forms passage.

3. bio-reactor according to claim 2, it is characterized in that, body outside screw (3) top is provided with corresponding projection one (9) place above one group of parallel projection one (9), pipe lid internal thread and is provided with one group of parallel projection two (10), when pipe lid (2) and body (1) rotate screw up time, projection one (9) and projection two (10) can engage mutually.

4. bio-reactor according to claim 3, is characterized in that, the number of described projection one (9), projection two (10) is between 1 ~ 5.